Chemical compounds

ABSTRACT

The specification relates to compounds of Formula (I): 
     
       
         
         
             
             
         
       
     
     and to pharmaceutically acceptable salts thereof, to processes and intermediates used for their preparation, to pharmaceutical compositions containing them and to their use in the treatment of cell proliferative disorders.

The specification relates to certain indazole compounds andpharmaceutically acceptable salts thereof that selectively down-regulatethe estrogen receptor and possess anti-cancer activity. Thespecification also relates to use of said indazole compounds andpharmaceutically acceptable salts thereof in methods of treatment of thehuman or animal body, for example in prevention or treatment of cancer.The specification also relates to processes and intermediate compoundsinvolved in the preparation of said indazole compounds and topharmaceutical compositions containing them.

Estrogen receptor alpha (ERα, ESR1, NR3A) and estrogen receptor beta(ERβ, ESR2, NR3b) are steroid hormone receptors which are members of thelarge nuclear receptor family. Structured similarly to all nuclearreceptors, ERα is composed of six functional domains (named A-F)(Dahlman-Wright, et al., Pharmacol. Rev., 2006, 58:773-781) and isclassified as a ligand-dependent transcription factor because after itsassociation with the specific ligand, (the female sex steroid hormone17b estradiol (E2)), the complex binds to genomic sequences, namedEstrogen Receptor Elements (ERE) and interacts with co-regulators tomodulate the transcription of target genes. The ERα gene is located on6q25.1 and encodes a 595AA protein and multiple isoforms can be produceddue to alternative splicing and translational start sites. In additionto the DNA binding domain (Domain C) and the ligand binding domain(Domain E) the receptor contains a N-terminal (A/B) domain, a hinge (D)domain that links the C and E domains and a C-terminal extension (Fdomain). While the C and E domains of ERα and ERβ are quite conserved(96% and 55% amino acid identity respectively) conservation of the A/B,D and F domains is poor (below 30% amino acid identity). Both receptorsare involved in the regulation and development of the femalereproductive tract and in addition play roles in the central nervoussystem, cardiovascular system and in bone metabolism. The genomic actionof ERs occurs in the nucleus of the cell when the receptor binds EREsdirectly (direct activation or classical pathway) or indirectly(indirect activation or non-classical pathway). In the absence ofligand, ERs are associated with heat shock proteins, Hsp90 and Hsp70,and the associated chaperone machinery stabilizes the ligand bindingdomain (LBD) making it accessible to ligand. Liganded ER dissociatesfrom the heat shock proteins leading to a conformational change in thereceptor that allows dimerisation, DNA binding, interaction withco-activators or co-repressors and modulation of target gene expression.In the non-classical pathway, AP-1 and Sp-1 are alternative regulatoryDNA sequences used by both isoforms of the receptor to modulate geneexpression. In this example, ER does not interact directly with DNA butthrough associations with other DNA bound transcription factors e.g.c-Jun or c-Fos (Kushner et al., Pure Applied Chemistry 2003,75:1757-1769). The precise mechanism whereby ER affects genetranscription is poorly understood but appears to be mediated bynumerous nuclear factors that are recruited by the DNA bound receptor.The recruitment of co-regulators is primarily mediated by two proteinsurfaces, AF2 and AF1 which are located in E-domain and the A/B domainrespectively. AF1 is regulated by growth factors and its activitydepends on the cellular and promoter environment whereas AF2 is entirelydependent on ligand binding for activity. Although the two domains canact independently, maximal ER transcriptional activity is achievedthrough synergistic interactions via the two domains (Tzukerman, et al.,Mol. Endocrinology, 1994, 8:21-30). Although ERs are consideredtranscription factors they can also act through non-genomic mechanismsas evidenced by rapid ER effects in tissues following E2 administrationin a timescale that is considered too fast for a genomic action. It isstill unclear if receptors responsible for the rapid actions of estrogenare the same nuclear ERs or distinct G-protein coupled steroid receptors(Warner, et al., Steroids 2006 71:91-95) but an increasing number of E2induced pathways have been identified e.g. MAPK/ERK pathway andactivation of endothelial nitric oxide synthase and PI3K/Akt pathway. Inaddition to ligand dependent pathways, ERα has been shown to have ligandindependent activity through AF-1 which has been associated withstimulation of MAPK through growth factor signalling e.g. insulin likegrowth factor 1 (IGF-1) and epidermal growth factor (EGF). Activity ofAF-1 is dependent on phosphorylation of Ser118 and an example ofcross-talk between ER and growth factor signalling is thephosphorylation of Ser 118 by MAPK in response to growth factors such asIGF-1 and EGF (Kato, et al., Science, 1995, 270:1491-1494).

A large number of structurally distinct compounds have been shown tobind to ER. Some compounds such as endogenous ligand E2, act as receptoragonists whereas others competitively inhibit E2 binding and act asreceptor antagonists. These compounds can be divided into 2 classesdepending on their functional effects. Selective estrogen receptormodulators (SERMs) such as tamoxifen have the ability to act as bothreceptor agonists and antagonists depending on the cellular and promotercontext as well as the ER isoform targeted. For example tamoxifen actsas an antagonist in breast but acts as a partial agonist in bone, thecardiovascular system and uterus. All SERMs appear to act as AF2antagonists and derive their partial agonist characteristics throughAF1. A second group, fulvestrant being an example, are classified asfull antagonists and are capable of blocking estrogen activity via thecomplete inhibition of AF1 and AF2 domains through induction of a uniqueconformation change in the ligand binding domain (LBD) on compoundbinding which results in complete abrogation of the interaction betweenhelix 12 and the remainder of the LBD, blocking co-factor recruitment(Wakeling, et al., Cancer Res., 1991, 51:3867-3873; Pike, et al.,Structure, 2001, 9:145-153).

Intracellular levels of ERα are down-regulated in the presence of E2through the ubiquitin/proteosome (Ub/26S) pathway. Polyubiquitinylationof liganded ERα is catalysed by at least three enzymes; theubiquitin-activating enzyme E1 activated ubiquitin is conjugated by E2with lysine residues through an isopeptide bond by E3 ubiquitin ligaseand polyubiquitinated ERα is then directed to the proteosome fordegradation. Although ER-dependent transcription regulation andproteosome-mediated degradation of ER are linked (Lonard, et al., Mol.Cell, 2000 5:939-948), transcription in itself is not required for ERαdegradation and assembly of the transcription initiation complex issufficient to target ERα for nuclear proteosomal degradation. This E2induced degradation process is believed to necessary for its ability torapidly activate transcription in response to requirements for cellproliferation, differentiation and metabolism (Stenoien, et al., Mol.Cell Biol., 2001, 21:4404-4412). Fulvestrant is also classified as aselective estrogen receptor down-regulator (SERD), a subset ofantagonists that can also induce rapid down-regulation of ERα via the26S proteosomal pathway. In contrast a SERM such as tamoxifen canincrease ERα levels although the effect on transcription is similar tothat seen for a SERD.

Approximately 70% of breast cancers express ER and/or progesteronereceptors implying the hormone dependence of these tumour cells forgrowth. Other cancers such as ovarian and endometrial are also thoughtto be dependent on ERα signalling for growth. Therapies for suchpatients can inhibit ER signalling either by antagonising ligand bindingto ER e.g. tamoxifen which is used to treat early and advanced ERpositive breast cancer in both pre and post menopausal setting;antagonising and down-regulating ERα e.g. fulvestrant which is used totreat breast cancer in women which have progressed despite therapy withtamoxifen or aromatase inhibitors; or blocking estrogen synthesis e.g.aromatase inhibitors which are used to treat early and advanced ERpositive breast cancer. Although these therapies have had an enormouslypositive impact on breast cancer treatment, a considerable number ofpatients whose tumours express ER display de novo resistance to existingER therapies or develop resistance to these therapies over time. Severaldistinct mechanisms have been described to explain resistance tofirst-time tamoxifen therapy which mainly involve the switch fromtamoxifen acting as an antagonist to an agonist, either through thelower affinity of certain co-factors binding to the tamoxifen-ERαcomplex being off-set by over-expression of these co-factors, or throughthe formation of secondary sites that facilitate the interaction of thetamoxifen-ERα complex with co-factors that normally do not bind to thecomplex. Resistance could therefore arise as a result of the outgrowthof cells expressing specific co-factors that drive the tamoxifen-ERαactivity. There is also the possibility that other growth factorsignalling pathways directly activate the ER receptor or co-activatorsto drive cell proliferation independently of ligand signalling.

More recently, mutations in ESR1 have been identified as a possibleresistance mechanism in metastatic ER-positive patient derived tumoursamples and patient-derived xenograft models (PDX) at frequenciesvarying from 17-25%. These mutations are predominantly, but notexclusively, in the ligand-binding domain leading to mutated functionalproteins; examples of the amino acid changes include Ser463Pro,Va1543Glu, Leu536Arg, Tyr537Ser, Tyr537Asn and Asp538Gly, with changesat amino acid 537 and 538 constituting the majority of the changescurrently described. These mutations have been undetected previously inthe genomes from primary breast samples characterised in the CancerGenome Atlas database. Of 390 primary breast cancer samples positive forER expression not a single mutation was detected in ESR1 (Cancer GenomeAtlas Network, 2012 Nature 490: 61-70). The ligand binding domainmutations are thought to have developed as a resistance response toaromatase inhibitor endocrine therapies as these mutant receptors showbasal transcriptional activity in the absence of estradiol. The crystalstructure of ER, mutated at amino acids 537 and 538, showed that bothmutants favoured the agonist conformation of ER by shifting the positionof helix 12 to allow co-activator recruitment and thereby mimickingagonist activated wild type ER. Published data has shown that endocrinetherapies such as tamoxifen and fulvestrant can still bind to ER mutantand inhibit transcriptional activation to some extent and thatfulvestrant is capable of degrading Try537Ser but that higher doses maybe needed for full receptor inhibition (Toy et al., Nat. Genetics 2013,45: 1439-1445; Robinson et al., Nat. Genetics 2013, 45: 144601451; Li,S. et al. Cell Rep. 4, 1116-1130 (2013). It is therefore feasible thatcertain compounds of the Formula (I) or pharmaceutically acceptablesalts thereof (as described hereinafter) will be capable ofdown-regulating and antagonising mutant ER although it is not known atthis stage whether ESR1 mutations are associated with an alteredclinical outcome.

Regardless of which resistance mechanism or combination of mechanismstakes place, many are still reliant on ER-dependent activities andremoval of the receptor through a SERD mechanism offers the best way ofremoving the ERα receptor from the cell. Fulvestrant is currently theonly SERD approved for clinical use, yet despite its mechanisticproperties, the pharmacological properties of the drug have limited itsefficacy due to the current limitation of a 500 mg monthly dose whichresults in less than 50% turnover of the receptor in patient samplescompared to the complete down-regulation of the receptor seen in invitro breast cell line experiments (Wardell, et al., Biochem. Pharm.,2011, 82:122-130). Hence there is a need for new ER targeting agentsthat have the required pharmaceutical properties and SERD mechanism toprovide enhanced benefit in the early, metastatic and acquiredresistance setting.

The compounds of the specification have been found to possess potentanti-tumour activity, being useful in inhibiting the uncontrolledcellular proliferation which arises from malignant disease. Thecompounds of the specification provide an anti-tumour effect by, as aminimum, acting as SERDs. For example, the compounds of thespecification may exhibit anti-tumour activity via the ability todown-regulate the estrogen receptor in a number of different breastcancer cell-lines, for example against the MCF-7, CAMA-1, BT474 and/orMDA-MB-134 breast cancer cell-lines. Such compounds may be expected tobe more suitable as therapeutic agents, particularly for the treatmentof cancer.

The compounds of the specification may also exhibit advantageousphysical properties (for example, lower lipophilicity, higher aqueoussolubility, higher permeability, lower plasma protein binding, and/orgreater chemical stability), and/or favourable toxicity profiles (forexample a decreased activity at hERG), and/or favourable metabolic orpharmacokinetic profiles, in comparison with other known SERDs. Suchcompounds may therefore be especially suitable as therapeutic agents,particularly for the treatment of cancer.

According to one aspect of the specification there is provided acompound of Formula (I):

wherein:

A is CR² or N; G is CR³ or N; D is CR⁴ or N; E is CR⁵ or N; Q is O, NHor NMe;

R¹ is CH₂F, CHF₂ or CF₃;

R² is H, F, Cl, Me, CN, OMe or OEt; R³ is H or F; R⁴ is H, F, CN or OMe;R⁵ is H or F;

R⁶ is H, Me, CH₂F, CHF₂ or CF₃;

R⁷ is H or Me;

R⁸ is C₁₋₃ alkyl, CH₂F, CHF₂, CF₃ or C₃₋₄ cycloalkyl;R⁹ is Me, F or CH₂F;R¹⁰ is Me, F, CH₂F, CHF₂, CF₃, CH₂OMe or CH₂OH;

R¹¹ is H or F; or

R¹⁰ and R¹¹ taken together with the carbon atom to which they areattached form a cyclopropyl ring or an oxetane ring;R¹² is independently selected from F or Me;

R¹³ is H or F; and

a is 0, 1 or 2;or a pharmaceutically acceptable salt thereof.

This specification also describes pharmaceutical compositions whichcomprise a compound of Formula (I), or a pharmaceutically acceptablesalt thereof, in association with a pharmaceutically acceptableexcipient.

This specification also describes a compound of Formula (I), or apharmaceutically acceptable salt thereof, for use as a medicament.

This specification also describes a compound of Formula (I), or apharmaceutically acceptable salt thereof, for use in the treatment ofcancer.

This specification also describes combinations of a compound of Formula(I), or a pharmaceutically acceptable salt thereof, with anotheranti-tumour agent, for use in the treatment of cancer.

Further aspects of the specification will be apparent to one skilled inthe art from reading this specification.

The C₁₋₃ alkyl group may be branched or unbranched. Examples of suitableC₁₋₃ alkyl groups are methyl (Me), ethyl (Et), n-propyl (n-Pr) ori-propyl (i-Pr).

The C₃₋₄ cycloalkyl group is an unsubstituted, monocyclic, unsaturatedcarbocyclic ring. Examples of suitable C₃₋₄ cycloalkyl groups arecyclopropyl and cyclobutyl.

In one embodiment there is provided a compound of Formula (I) as definedabove.

In one embodiment there is provided a pharmaceutically acceptable saltof a compound of Formula (I).

In one embodiment A is CR².

In one embodiment G is CR³.

In one embodiment A is CR² and G is CR³.

In one embodiment A is CR² and G is N.

In one embodiment A is N and G is CR³.

In one embodiment R² is H, F, Cl, Me, CN or OMe.

In one embodiment R² is H, F or OMe.

In one embodiment R² is H or F.

In one embodiment R² is H.

In one embodiment R² is F.

In one embodiment A is CR² and R² is H, F or OMe.

In one embodiment G is CR³ and R³ is H, F or OMe.

In one embodiment A is CH and G is CH.

In one embodiment A is C—F and G is C—F.

In one embodiment A is C—F and G is CH.

In one embodiment A is C—OMe and G is CH.

In one embodiment A is C—OMe and G is C—F.

In one embodiment one of A or G is CH and the other of A or G is N.

In one embodiment D is CR⁴.

In one embodiment E is CR⁵.

In one embodiment both D and E are CH.

In one embodiment both D and E are N.

In one embodiment one of D or E is CH and the other of D or E is N.

In one embodiment one of D or E is C—F and the other of D or E is CH.

In one embodiment one of D or E is C—OMe and the other of D or E is CH.

In one embodiment Q is O or NH.

In one embodiment Q is O.

In one embodiment Q is NH.

In one embodiment Q is NMe.

In one embodiment R¹ is CH₂F or CHF₂.

In one embodiment R¹ is CH₂F.

In one embodiment R¹ is CHF₂.

In one embodiment R¹ is CF₃.

In one embodiment R⁶ is H or Me.

In one embodiment R⁶ is H.

In one embodiment R⁶ is Me.

In one embodiment R⁷ is H.

In one embodiment R⁷ is Me.

In one embodiment R⁸ is C₁₋₃ alkyl, CHF₂ or cyclopropyl.

In one embodiment R⁸ is C₁₋₃ alkyl or CHF₂.

In one embodiment R⁸ is C₁₋₃ alkyl.

In one embodiment R⁸ is methyl.

In one embodiment R⁸ is CHF₂.

In one embodiment R⁹ is Me or F.

In one embodiment R⁹ is Me.

In one embodiment R⁹ is F.

In one embodiment R¹⁰ is Me, F, CH₂F, CH₂OMe or CH₂OH.

In one embodiment R¹⁰ is Me, F, CH₂OMe or CH₂OH.

In one embodiment R¹⁰ is F, CH₂OMe or CH₂OH.

In one embodiment R¹⁰ is CH₂OMe or CH₂OH.

In one embodiment R¹⁰ is F.

In one embodiment R¹ is F.

In one embodiment R¹¹ is H.

In one embodiment R¹⁰ and R¹¹ taken together with the carbon atom towhich they are attached form a cyclopropyl ring or an oxetane ring.

In one embodiment R¹⁰ and R¹¹ taken together with the carbon atom towhich they are attached form a cyclopropyl ring.

In one embodiment R¹⁰ and R¹¹ taken together with the carbon atom towhich they are attached form an oxetane ring.

In one embodiment R¹⁰ is CH₂OMe or CH₂OH and R¹¹ is F.

In one embodiment R¹⁰ is CH₂OMe or CH₂OH and R⁹ is Me.

In one embodiment R¹⁰ is F and R⁹ is F.

In one embodiment R⁹ is Me or F and R² is H, F, Cl, Me, CN or OMe.

In one embodiment R⁹ is Me and R¹¹ is F.

In one embodiment R⁹ is F and R¹¹ is Me.

In one embodiment R⁹ is F and R¹¹ is F.

In one embodiment R⁹ is F and R¹⁰ and R¹¹ taken together with the carbonatom to which they are attached form a cyclopropyl ring or an oxetanering.

In one embodiment R⁹ is F and R¹⁰ and R¹¹ taken together with the carbonatom to which they are attached form a cyclopropyl ring.

In one embodiment R⁹ is F and R¹⁰ and R¹¹ taken together with the carbonatom to which they are attached form an oxetane ring.

In one embodiment the group —CH₂—C(R⁹)(R¹⁰)(R¹¹) in the compound ofFormula (I) is selected from the group consisting of:

In one embodiment the group —CH₂—C(R⁹)(R¹⁰)(R¹¹) in the compound ofFormula (I) is selected from the group consisting of:

In one embodiment the group —CH₂—C(R⁹)(R¹⁰)(R¹¹) in the compound ofFormula (I) is selected from the group consisting of:

In one embodiment the group —CH₂—C(R⁹)(R¹⁰)(R¹¹) in the compound ofFormula (I) is selected from the group consisting of:

In one embodiment the group —CH₂—C(R⁹)(R¹⁰)(R¹¹) in the compound ofFormula (I) is selected from the group consisting of:

In one embodiment the group —CH₂—C(R⁹)(R¹⁰)(R¹¹) in the compound ofFormula (I) is selected from the group consisting of:

In one embodiment R¹² is Me.

In one embodiment R¹² is F.

In one embodiment R¹³ is H.

In one embodiment R¹³ is F.

In one embodiment a is 0 or 1.

In one embodiment a is 0.

In one embodiment a is 1.

In one embodiment a is 2.

In one embodiment there is provided a compound of Formula (IA):

wherein:

Q is O, NH or NMe;

R¹ is CH₂F, CHF₂ or CF₃;

R⁶ is H or Me; R⁷ is H or Me;

R⁸ is Me, CHF₂ or cyclopropyl;

R⁹ is Me or F;

R¹⁰ is Me, F, CH₂F, CH₂OMe or CH₂OH;

R¹¹ is H or F; or

R¹⁰ and R¹¹ taken together with the carbon atom to which they areattached form a cyclopropyl ring or an oxetane ring;R¹² is independently selected from H or Me;

R¹³ is H or F;

a is 0, 1 or 2; andRing Y is selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.

In one embodiment there is provided a compound of Formula (IA), or apharmaceutically acceptable salt thereof, wherein Ring Y is selectedfrom the group consisting of:

In one embodiment there is provided a compound of Formula (IA), or apharmaceutically acceptable salt thereof, wherein Ring Y is selectedfrom the group consisting of:

In one embodiment there is provided a compound of Formula (IA), or apharmaceutically acceptable salt thereof, wherein Ring Y is selectedfrom the group consisting of:

In one embodiment there is provided a compound of Formula (IA), or apharmaceutically acceptable salt thereof, wherein Ring Y is selectedfrom the group consisting of:

In one embodiment there is provided a compound of Formula (IA), or apharmaceutically acceptable salt thereof, wherein Ring Y is selectedfrom the group consisting of:

In one embodiment there is provided a compound of Formula (IA), or apharmaceutically acceptable salt thereof, wherein Q is NH.

In one embodiment there is provided a compound of Formula (IA), or apharmaceutically acceptable salt thereof, wherein Q is O.

In one embodiment there is provided a compound of Formula (IA), or apharmaceutically acceptable salt thereof, wherein R¹ is CH₂F.

In one embodiment there is provided a compound of Formula (IA), or apharmaceutically acceptable salt thereof, wherein R¹ is CHF₂.

In one embodiment there is provided a compound of Formula (IA), or apharmaceutically acceptable salt thereof, wherein R⁶ is H.

In one embodiment there is provided a compound of Formula (IA), or apharmaceutically acceptable salt thereof, wherein R⁷ is H.

In one embodiment there is provided a compound of Formula (IA), or apharmaceutically acceptable salt thereof, wherein R⁸ is Me.

In one embodiment there is provided a compound of Formula (IA), or apharmaceutically acceptable salt thereof, wherein R¹⁰ is F, CH₂OMe orCH₂OH.

In one embodiment there is provided a compound of Formula (IA), or apharmaceutically acceptable salt thereof, wherein R⁹ is F and R¹⁰ andR¹¹ taken together with the carbon atom to which they are attached forma cyclopropyl ring or an oxetane ring. In a further embodiment R¹⁰ andR¹¹ taken together with the carbon atom to which they are attached forma cyclopropyl ring. In a further embodiment R¹⁰ and R¹¹ taken togetherwith the carbon atom to which they are attached form an oxetane ring.

In one embodiment there is provided a compound of Formula (IA), or apharmaceutically acceptable salt thereof, wherein R⁸ is Me.

In one embodiment the group —CH₂—C(R⁹)(R¹⁰)(R¹¹) in the compound ofFormula (IA) is selected from the group consisting of:

In one embodiment the group —CH₂—C(R⁹)(R¹⁰)(R¹¹) in the compound ofFormula (IA) is selected from the group consisting of:

In one embodiment the group —CH₂—C(R⁹)(R¹⁰)(R¹¹) in the compound ofFormula (IA) is selected from the group consisting of:

In one embodiment the group —CH₂—C(R⁹)(R¹⁰)(R¹¹) in the compound ofFormula (IA) is selected from the group consisting of:

In one embodiment there is provided a compound of Formula (IA), or apharmaceutically acceptable salt thereof, wherein R¹³ is H.

In one embodiment there is provided a compound of Formula (IA), or apharmaceutically acceptable salt thereof, wherein a is 0.

In one embodiment there is provided a compound of Formula (IB):

wherein:

Q is O, NH or NMe;

R¹ is CH₂F, CHF₂ or CF₃;

R⁶ is H or Me, R⁷ is H or Me;

R⁸ is Me, CHF₂ or cyclopropyl;R¹⁴ is selected from the group consisting of:

and Ring Y is selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.

In one embodiment there is provided a compound of Formula (IB), or apharmaceutically acceptable salt thereof, wherein Ring Y is selectedfrom the group consisting of:

In one embodiment there is provided a compound of Formula (IB), or apharmaceutically acceptable salt thereof, wherein Ring Y is selectedfrom the group consisting of:

In one embodiment there is provided a compound of Formula (IB), or apharmaceutically acceptable salt thereof, wherein Ring Y is selectedfrom the group consisting of:

In one embodiment there is provided a compound of Formula (IB), or apharmaceutically acceptable salt thereof, wherein Ring Y is selectedfrom the group consisting of:

In one embodiment there is provided a compound of Formula (IB), or apharmaceutically acceptable salt thereof, wherein Ring Y is selectedfrom the group consisting of:

In one embodiment there is provided a compound of Formula (IB), or apharmaceutically acceptable salt thereof, wherein Q is NH.

In one embodiment there is provided a compound of Formula (IB), or apharmaceutically acceptable salt thereof, wherein Q is O.

In one embodiment there is provided a compound of Formula (IB), or apharmaceutically acceptable salt thereof, wherein R¹ is CH₂F.

In one embodiment there is provided a compound of Formula (IB), or apharmaceutically acceptable salt thereof, wherein R¹ is CHF₂.

In one embodiment there is provided a compound of Formula (IB), or apharmaceutically acceptable salt thereof, wherein R⁶ is H.

In one embodiment there is provided a compound of Formula (IB), or apharmaceutically acceptable salt thereof, wherein R⁷ is H.

In one embodiment there is provided a compound of Formula (IB), or apharmaceutically acceptable salt thereof, wherein R⁸ is Me.

In one embodiment the group R¹⁴ in the compound of Formula (IB) isselected from the group consisting of:

In one embodiment the group R¹⁴ in the compound of Formula (IB) isselected from the group consisting of:

In one embodiment the group R¹⁴ in the compound of Formula (IB) isselected from the group consisting of:

In a further aspect of the specification there is provided the compoundof Formula (IC):

wherein:

A is CR² or N; s G is CR³ or N; D is CR⁴ or N; E is CR⁵ or N; Q is O, NHor NMe;

R¹ is CH₂F, CHF₂ or CF₃;

R² is H, F, Cl, Me, CN, OMe or OEt; R³ is H or F; R⁴ is H, F, CN or OMe;R⁵ is H or F;

R⁶ is H, Me, CH₂F, CHF₂ or CF₃;

R⁷ is H or Me;

R⁸ is C₁₋₃ alkyl, CH₂F, CHF₂, CF₃ or C₃₋₄ cycloalkyl;R⁹ is Me, F or CH₂F;R¹⁰ is Me, F, CH₂F, CHF₂, CF₃, CH₂OMe or CH₂OH;

R¹¹ is H or F; or

R¹⁰ and R¹¹ taken together with the carbon atom to which they areattached form a cyclopropyl ring or an oxetane ring;R¹² is independently selected from F or Me;

R¹³ is H or F; and

a is 0, 1 or 2;or a pharmaceutically acceptable salt thereof.

In one embodiment there is provided the compound of Formula (IC) or apharmaceutically acceptable salt thereof wherein R² is H, F, Cl, Me, CNor OMe and R⁹ is Me or F.

In a further embodiment there is provided the compound of Formula (IC)or a pharmaceutically acceptable salt thereof wherein thestereochemistry at the 6-position of the pyrazolo[4,3-f]isoquinolinering is S.

In a further embodiment there is provided the compound of Formula (IC)or a pharmaceutically acceptable salt thereof wherein thestereochemistry at the 6-position of the pyrazolo[4,3-f]isoquinolinering is R.

In a further embodiment there is provided the compound of Formula (IC)or a pharmaceutically acceptable salt thereof wherein thestereochemistry at the 8-position of the pyrazolo[4,3-f]isoquinolinering is S.

In a further embodiment there is provided the compound of Formula (IC)or a pharmaceutically acceptable salt thereof wherein thestereochemistry at the 8-position of the pyrazolo[4,3-f]isoquinolinering is R.

In one embodiment there is provided a compound of Formula (ID):

wherein:

Q is O or NH;

R¹ is CH₂F or CHF₂;R¹⁴ is selected from the group consisting of:

and Ring Y is selected from the group consisting of:

In one embodiment there is provided a compound of Formula (ID), or apharmaceutically acceptable salt thereof, wherein Q is NH.

In one embodiment there is provided a compound of Formula (ID), or apharmaceutically acceptable salt thereof, wherein Q is O.

In one embodiment there is provided a compound of Formula (ID), or apharmaceutically acceptable salt thereof, wherein R¹ is CH₂F.

In one embodiment there is provided a compound of Formula (ID), or apharmaceutically acceptable salt thereof, wherein Q is O and R¹ is CH₂F.

In one embodiment there is provided a compound of Formula (ID), or apharmaceutically acceptable salt thereof, wherein Q is NH and R¹ isCH₂F.

In one embodiment there is provided a compound of Formula (ID), or apharmaceutically acceptable salt thereof, wherein R¹⁴ is selected fromthe group consisting of:

In one embodiment there is provided a compound of Formula (ID), or apharmaceutically acceptable salt thereof, wherein Ring Y is selectedfrom the group consisting of:

In one embodiment there is provided a compound of Formula (I), whereinthe compound is selected from the group consisting of:

-   N-(4-((6S,8R)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   6-((6S,8R)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;-   6-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;-   N-(4-((6S,8R)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   3-((6S,8R)-6-(2,6-difluoro-4-((1-(3-fluoropropyl)azetidin-3-yl)amino)phenyl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)-2,2-difluoropropan-1-ol;-   N-(4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   (6S,8R)-7-((1-fluorocyclopropyl)methyl)-6-(4-((1-(3-fluoropropyl)azetidin-3-yl)oxy)-2-methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline;-   N-(3,5-difluoro-4-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine;    and-   5-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)-4-methoxypyridin-2-amine;    or a pharmaceutically acceptable salt thereof.

In one embodiment there is provided a compound of Formula (I), whereinthe compound is selected from the group consisting of:

-   N-(4-((6S,8R)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   6-((6S,8R)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;-   6-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;-   N-(4-((6S,8R)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   3-((6S,8R)-6-(2,6-difluoro-4-((1-(3-fluoropropyl)azetidin-3-yl)amino)phenyl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)-2,2-difluoropropan-1-ol;-   N-(4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   (6S,8R)-7-((1-fluorocyclopropyl)methyl)-6-(4-((1-(3-fluoropropyl)azetidin-3-yl)oxy)-2-methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline;-   N-(3,5-difluoro-4-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   5-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)-4-methoxypyridin-2-amine;-   N-(4-((6S,8R)-7-((3-(fluoromethyl)oxetan-3-yl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   N-(3,5-difluoro-4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   (6S,8R)-7-(2-fluoro-3-methoxy-2-methylpropyl)-6-(4-(1-(3-fluoropropyl)azetidin-3-yloxy)-2-methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline;-   N-(4-((6S,8R)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   2,2-difluoro-3-((6S,8R)-6-(5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol;-   N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine;-   5-fluoro-6-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;-   N-(4-((6S,8R)-7-(2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   N-(3-ethoxy-4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   N-(4-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   (6S,8R)-7-(2,2-difluoroethyl)-6-(4-((1-(3-fluoropropyl)azetidin-3-yl)oxy)-2-methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline;-   3-((6S,8R)-6-(2,6-difluoro-4-(1-(3-fluoropropyl)azetidin-3-ylamino)phenyl)-8-methyl-8,9-dihydro-3H-pyrazolo[4,3-f]isoquinolin-7(6H)-yl)-2-fluoro-2-methylpropan-1-ol;-   6-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;    and-   (6S,8R)-7-(2,2-difluoroethyl)-6-(5-((1-(3-fluoropropyl)azetidin-3-yl)oxy)pyridin-2-yl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline;    or a pharmaceutically acceptable salt thereof.

In one embodiment there is provided a compound of Formula (I), whereinthe compound is selected from the group consisting of:

-   1-(3-fluoropropyl)-N-(4-((6R,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-amine;-   1-(3-fluoropropyl)-N-(3-methoxy-4-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-amine;-   2-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine;-   5-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine;-   2,2-difluoro-3-((6S,8R)-6-(3-fluoro-5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol;-   6-((6S,8R)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,9,8-tetrahydro-3H-pyrazolo[4,3-f]isoquin-6-yl)-N-(3-fluoropropyl)azetidi-3-yl)pyridin-3-amine;-   5-fluoro-6-((6S,8R)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;-   N-(1-(3-fluoropropyl)azetidin-3-yl)-N-methyl-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine;-   N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-6-deuterio-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine;-   N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-1-deuterio-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine;-   N-(1-(3-fluoropropyl)azetidin-3-yl)-5-methoxy-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine;-   5-((6S,8R)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyrazin-2-amine;-   2-fluoro-6-((6S,8R)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;-   6-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)-5-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-2-amine;-   N-(2-fluoro-4-((6R,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   N-(1-(3-fluoropropyl)azetidin-3-yl)-5-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyrazin-2-amine;-   6-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)-N-methylpyridin-3-amine;-   6-((6S,8R)-7-(2,2-difluoroethyl)-6,8-dimethyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;-   is    N-(4-((6S,8R)-7-(2,2-difluoroethyl)-6,8-dimethyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   6-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-5-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;-   6-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-2-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;-   N-(4-((6R,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   N-(4-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   5-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyrazin-2-amine;-   6-((6S,8S)-7-(2,2-difluoroethyl)-8-(difluoromethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;-   N-(4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)-N-methylazetidin-3-amine;-   N-(2-fluoro-4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-5-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   5-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-2-amine;-   5-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyrazin-2-amine;-   N-(4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3,3,3-trifluoropropyl)azetidin-3-amine;-   6-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)-N-methylpyridin-3-amine;-   2,2-difluoro-3-((6S,8R)-6-(5-((1-(3-fluoropropyl)azetidin-3-yl)oxy)pyridin-2-yl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol;-   2,2-difluoro-3-((6S,8R)-6-(3-fluoro-5-((1-(3-fluoropropyl)azetidin-3-yl)oxy)pyridin-2-yl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol;-   2,2-difluoro-3-((6S,8R)-6-(4-((1-(3-fluoropropyl)azetidin-3-yl)amino)-2-methoxyphenyl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol;-   2,2-difluoro-3-((6S,8R)-1-fluoro-6-(5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol;-   2,2-difluoro-3-((6S,8R)-1-fluoro-6-(6-fluoro-5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol;-   6-((6S,8R)-7-(2-fluoro-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;-   N-(4-((6S,8R)-7-(2-fluoro-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;-   6-((6S,8R)-7-(2-fluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;-   6-((6S,8R)-7-(2-fluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;-   1-(3-fluoropropyl)-N-(4-((6S,8R)-7-(2-fluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)azetidin-3-amine;-   1-(3-fluoropropyl)-N-(4-((6S,8R)-7-(2-fluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)azetidin-3-amine;-   N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-7-isobutyl-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine;-   6-((6S,8R)-7-(2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;-   5-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,3-trifluoropropyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine;-   (S)-6-(8,8-dimethyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;    and-   (S)-6-(7-(2,2-difluoroethyl)-8,8-dimethyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine    or a pharmaceutically acceptable salt thereof.-   In one embodiment there is provided a compound of Formula (I) which    is    N-(1-(3-fluoropropyl)azetidin-3-yl)-6-(8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine,    or a pharmaceutically acceptable salt thereof.

In one embodiment there is provided a compound of Formula (I) which isselected from the group consisting of:

-   N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine;    and-   N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6R,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine;    or a pharmaceutically acceptable salt thereof.

In one embodiment there is provided a compound of Formula (I) which isN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine,or a pharmaceutically acceptable salt thereof.

In one embodiment there is provided a compound of Formula (I) which isN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6R,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine,or a pharmaceutically acceptable salt thereof.

In one embodiment there is provided a compound of Formula (I), or apharmaceutically acceptable salt thereof, wherein the compound isselected from any of the Examples in the specification. A furtherfeature is any of the embodiments described in the specification withthe proviso that any of the specific Examples are individuallydisclaimed. A further feature is any of the embodiments described in thespecification with the proviso that any one or more of the compoundsselected from the above list of examples of compounds of thespecification are individually disclaimed.

For the avoidance of doubt, when a is 1, the R¹² substituent may besubstituted on either carbon of the respective ethyl chain with which itis associated, and when a is 2, the R¹² substituent may be substitutedat either a single carbon or at both carbons of the said ethyl chain.When a is 1 or 2, therefore, the following substitution patterns arepossible, each of which represent a further embodiment:

In a further embodiment, therefore, a is 1 or 2 and R¹² is attached tothe remainder of the compound of Formula (I) as shown in (a) below:

In a further embodiment, a is 1 or 2, R¹² is methyl, and R¹² is attachedto the remainder of the compound of Formula (I) as shown in (b) below:

For the further avoidance of doubt, the use of “

” in formulas of this specification denotes the point of attachmentbetween different groups.

The compounds of Formula (I) have two or more chiral centres and it willbe recognised that the compound of Formula (I) may be prepared, isolatedand/or supplied with or without the presence, in addition, of one ormore of the other possible enantiomeric and/or diastereomeric isomers ofthe compound of Formula (I) in any relative proportions. The preparationof enantioenriched/enantiopure and/or diastereoenriched/diastereopurecompounds may be carried out by standard techniques of organic chemistrythat are well known in the art, for example by synthesis fromenantioenriched or enantiopure starting materials, use of an appropriateenantioenriched or enantiopure catalyst during synthesis, and/or byresolution of a racemic or partially enriched mixture of stereoisomers,for example via chiral chromatography.

For use in a pharmaceutical context it may be preferable to provide acompound of Formula (I) or a pharmaceutically acceptable salt thereofwithout large amounts of the other stereoisomeric forms being present.

Accordingly, in one embodiment there is provided a compositioncomprising a compound of Formula (I) or a pharmaceutically acceptablesalt thereof, optionally together with one or more of the otherstereoisomeric forms of the compound of Formula (I) or pharmaceuticallyacceptable salt thereof, wherein the compound of Formula (I) orpharmaceutically acceptable salt thereof is present within thecomposition with a diastereomeric excess (% de) of ≥90%.

In a further embodiment the % de in the above-mentioned composition is≥95%.

In a further embodiment the % de in the above-mentioned composition is≥98%.

In a further embodiment the % de in the above-mentioned composition is≥99%.

In a further embodiment there is provided a composition comprising acompound of Formula (I) or a pharmaceutically acceptable salt thereof,optionally together with one or more of the other stereoisomeric formsof the compound of Formula (I) or pharmaceutically acceptable saltthereof, wherein the compound of Formula (I) or pharmaceuticallyacceptable salt thereof is present within the composition with anenantiomeric excess (% ee) of ≥90%.

In a further embodiment the % ee in the above-mentioned composition is≥95%.

In a further embodiment the % ee in the above-mentioned composition is≥98%.

In a further embodiment the % ee in the above-mentioned composition is≥99%.

In a further embodiment there is provided a composition comprising acompound of Formula (I) or a pharmaceutically acceptable salt thereof,optionally together with one or more of the other stereoisomeric formsof the compound of Formula (I) or pharmaceutically acceptable saltthereof, wherein the compound of Formula (I) or pharmaceuticallyacceptable salt thereof is present within the composition with anenantiomeric excess (% ee) of ≥90% and a diastereomeric excess (% de) of≥90%.

In further embodiments of the above-mentioned composition the % ee and %de may take any combination of values as listed below:

-   -   The % ee is ≤5% and the % de is ≥80%.    -   The % ee is ≤5% and the % de is ≥90%.    -   The % ee is ≤5% and the % de is ≥95%.    -   The % ee is ≤5% and the % de is ≥98%.    -   The % ee is ≥95% and the % de is ≥95%.    -   The % ee is ≥98% and the % de is ≥98%.    -   The % ee is ≥99% and the % de is ≥99%.

In a further embodiment there is provided a pharmaceutical compositionwhich comprises a compound of the Formula (I) or a pharmaceuticallyacceptable salt thereof, in association with a pharmaceuticallyacceptable excipient.

In one embodiment there is provided a pharmaceutical composition whichcomprises a compound of the Formula (I) or a pharmaceutically acceptablesalt thereof, in association with a pharmaceutically acceptableexcipient, optionally further comprising one or more of the otherstereoisomeric forms of the compound of Formula (I) or pharmaceuticallyacceptable salt thereof, wherein the compound of Formula (I) orpharmaceutically acceptable salt thereof is present within thecomposition with an enantiomeric excess (% ee) of ≥90%.

In a further embodiment the % ee in the above-mentioned composition is≥95%.

In a further embodiment the % ee in the above-mentioned composition is≥98%.

In a further embodiment the % ee in the above-mentioned composition is≥99%.

In one embodiment there is provided a pharmaceutical composition whichcomprises a compound of the Formula (I) or a pharmaceutically acceptablesalt thereof, in association with a pharmaceutically acceptableexcipient, optionally further comprising one or more of the otherstereoisomeric forms of the compound of Formula (I) or pharmaceuticallyacceptable salt thereof, wherein the compound of Formula (I) orpharmaceutically acceptable salt thereof is present within thecomposition with a diastereomeric excess (% de) of ≥90%.

In a further embodiment the % de in the above-mentioned composition is≥95%.

In a further embodiment the % de in the above-mentioned composition is≥98%.

In a further embodiment the % de in the above-mentioned composition is≥99%.

In one embodiment there is provided a pharmaceutical composition whichcomprises a compound of the Formula (I) or a pharmaceutically acceptablesalt thereof, in association with a pharmaceutically acceptableexcipient, optionally further comprising one or more of the otherstereoisomeric forms of the compound of Formula (I) or pharmaceuticallyacceptable salt thereof, wherein the compound of Formula (I) orpharmaceutically acceptable salt thereof is present within thecomposition with an enantiomeric excess (% ee) of ≥90% and adiastereomeric excess (% de) of ≥90%.

In further embodiments of the above-mentioned pharmaceutical compositionthe % ee and % de may take any combination of values as listed below:

-   -   The % ee is ≥95% and the % de is ≥95%.    -   The % ee is ≥98% and the % de is ≥98%.    -   The % ee is ≥99% and the % de is ≥99%.

The compounds of Formula (I) and pharmaceutically acceptable saltsthereof may be prepared, used or supplied in amorphous form, crystallineform, or semicrystalline form and any given compound of Formula (I) orpharmaceutically acceptable salt thereof may be capable of being formedinto more than one crystalline/polymorphic form, including hydrated(e.g. hemi-hydrate, a mono-hydrate, a di-hydrate, a tri-hydrate or otherstoichiometry of hydrate) and/or solvated forms. It is to be understoodthat the present specification encompasses any and all such solid formsof the compound of Formula (I) and pharmaceutically acceptable saltsthereof.

In further embodiments there is provided a compound of Formula (I),which is obtainable by the methods described in the ‘Examples’ sectionhereinafter.

The present specification is intended to include all isotopes of atomsoccurring in the present compounds. Isotopes will be understood toinclude those atoms having the same atomic number but different massnumbers. For example, isotopes of hydrogen include tritium anddeuterium. Isotopes of carbon include ¹³C and ¹⁴C. Isotopes of nitrogeninclude ¹⁵N. In a particular embodiment there is provided a compound ofFormula (I) wherein R⁶ is deuterium.

A suitable pharmaceutically acceptable salt of a compound of the Formula(I) is, for example, an acid addition salt. A suitable pharmaceuticallyacceptable salt of a compound of Formula (I) may be, for example, anacid-addition salt of a compound of the Formula (I), for example anacid-addition salt with an inorganic or organic acid such as aceticacid, adipic acid, benzene sulfonic acid, benzoic acid, cinnamic acid,citric acid, D,L-lactic acid, ethane disulfonic acid, ethane sulfonicacid, fumaric acid, hydrochloric acid, L-tartaric acid, maleic acid,malic acid, malonic acid, methane sulfonic acid, napadisylic acid,phosphoric acid, saccharin, succinic acid, sulfuric acid,p-toluenesulfonic acid, toluene sulfonic acid or trifluoroacetic acid.

A further suitable pharmaceutically acceptable salt of a compound of theFormula (I) is, for example, a salt formed within the human or animalbody after administration of a compound of the Formula (I) to said humanor animal body.

The compound of Formula (I) or pharmaceutically acceptable salt thereofmay be prepared as a co-crystal solid form. It is to be understood thata pharmaceutically acceptable co-crystal of a compound of the Formula(I) or pharmaceutically acceptable salts thereof, form an aspect of thepresent specification.

The specific solid forms described herein provide X-ray powderdiffraction patterns substantially the same as the X-ray powderdiffraction patterns shown in the Figures and have the various 2-thetavalues as shown in the Tables included herein. It will be understoodthat the 2-theta values of an X-ray powder diffraction pattern may varyslightly from one machine to another or from one sample to another, andso the values quoted are not to be construed as absolute.

It is known that an X-ray powder diffraction pattern may be obtainedwhich has one or more measurement errors depending on measurementconditions (such as equipment or machine used). In particular, it isgenerally known that intensities in an X-ray powder diffraction patternmay fluctuate depending on measurement conditions. Therefore it shouldbe understood that the solid forms of the present specification are notlimited to the crystals that provide X-ray powder diffraction patternsthat are identical to the X-ray powder diffraction pattern shown in theFigures, and any crystals providing X-ray powder diffraction patternssubstantially the same as those shown in the Figures fall within thescope of the present specification. A person skilled in the art of X-raypowder diffraction is able to judge the substantial identity of X-raypowder diffraction patterns.

Persons skilled in the art of X-ray powder diffraction will realise thatthe relative intensity of peaks can be affected by, for example, grainsabove 30 μm in size and non-unitary aspect ratios, which may affectanalysis of samples. The skilled person will also realise that theposition of reflections can be affected by the precise height at whichthe sample sits in the diffractometer and the zero calibration of thediffractometer. The surface planarity of the sample may also have asmall effect. Hence the diffraction pattern data presented are not to betaken as absolute values. (Jenkins, R & Snyder, R. L. ‘Introduction toX-Ray Powder Diffractometry’ John Wiley & Sons 1996; Bunn, C. W. (1948),Chemical Crystallography, Clarendon Press, London; Klug, H. P. &Alexander, L. E. (1974), X-Ray Diffraction Procedures).

Generally, a measurement error of a diffraction angle in an X-ray powderdiffractogram is approximately plus or minus 0.2° 2-theta, and suchdegree of a measurement error should be taken into account whenconsidering the X-ray powder diffraction pattern in the Figures and whenreading data contained in the Tables included herein. Furthermore, itshould be understood that intensities might fluctuate depending onexperimental conditions and sample preparation (preferred orientation).

In this specification the form of the compoundN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine(hereafter Compound X) was initially found to be an amorphous solid.Useful crystalline polymorphic forms of the compound have subsequentlybeen produced using the conditions described in the experimentalsection.

Therefore in a further aspect of the specification there is providedpolymorphic Form A of Compound X. This polymorphic form may becharacterised in that it provides at least one of the following 2θvalues measured using CuKa radiation: 15.5, 18.6 and 24.6°.

Polymorphic Form A of Compound X is characterised in providing an X-raypowder diffraction pattern, substantially as shown in FIG. 1.

Ten X-Ray powder diffraction peaks for this polymorphic form [Angle2-theta (20), Intensity (%)] are: 21.1 (100%), 20.8 (54.3%), 14.6(41.9%), 18.6 (41.6%), 12.3 (38.9%), 15.5 (34.1%), 24.6 (31.3%), 15.8(30.6%), 13.4 (23.2%) and 19.0° (21.7%).

According to the present specification there is provided the polymorphicForm A of Compound X, which has an X-ray powder diffraction pattern withat least one specific peak at about 2-theta=15.5°.

According to the present specification there is provided the polymorphicForm A of Compound X, which has an X-ray powder diffraction pattern withat least one specific peak at about 2-theta=18.6°.

According to the present specification there is provided the polymorphicForm A of Compound X, which has an X-ray powder diffraction pattern withat least one specific peak at about 2-theta=24.6°.

According to the present specification there is provided the polymorphicForm A of Compound X, which has an X-ray powder diffraction pattern withat least two specific peaks at about 2-theta=15.5° and 18.6°.

According to the present specification there is provided the polymorphicForm A of Compound X, which has an X-ray powder diffraction pattern withspecific peaks at about 2-theta=21.1, 20.8, 14.6, 18.6, 12.3, 15.5,24.6, 15.8, 13.4 and 19.0°.

According to the present specification there is provided polymorphicForm A of Compound X which has an X-ray powder diffraction patternsubstantially the same as the X-ray powder diffraction pattern shown inFIG. 1.

According to the present specification there is provided polymorphicForm A of Compound X, which has an X-ray powder diffraction pattern withat least one specific peak at 2-theta=15.5° plus or minus 0.2° 2-theta.

According to the present specification there is provided a polymorphicForm A of Compound X, which has an X-ray powder diffraction pattern withat least one specific peak at 2-theta=18.6° plus or minus 0.2° 2-theta.

According to the present specification there is provided the polymorphicForm A of Compound X, which has an X-ray powder diffraction pattern withat least two specific peaks at 2-theta=15.5° and 18.6° wherein saidvalues may be plus or minus 0.2° 2-theta.

According to the present specification there is provided a polymorphicForm A of Compound X, which has an X-ray powder diffraction pattern withspecific peaks at 2-theta=21.1, 20.8, 14.6, 18.6, 12.3, 15.5, 24.6,15.8, 13.4 and 19.0° wherein said values may be plus or minus 0.2°2-theta.

In a further aspect of the specification there is provided polymorphicForm E of Compound X. This polymorphic form may be characterised in thatit provides at least one of the following 2θ values measured using CuKaradiation: 14.8, 16.2 and 17.9°.

Polymorphic Form E of Compound X is characterised in providing an X-raypowder diffraction pattern, substantially as shown in FIG. 8.

Ten X-Ray powder diffraction peaks for this polymorphic form [Angle2-theta (20), Intensity (%)] are: 17.9 (100%), 14.8 (67.1%), 20.9(60.1%), 23.1 (55.4%), 16.2 (49.3%), 20.0 (35.6%), 18.2 (32.9%), 12.3(30.4%), 22.2 (19.0%) and 13.9° (18.9%).

According to the present specification there is provided the polymorphicForm E of Compound X, which has an X-ray powder diffraction pattern withat least one specific peak at about 2-theta=17.9°.

According to the present specification there is provided the polymorphicForm E of Compound X, which has an X-ray powder diffraction pattern withat least one specific peak at about 2-theta=14.8°.

According to the present specification there is provided the polymorphicForm E of Compound X, which has an X-ray powder diffraction pattern withat least one specific peak at about 2-theta=17.9°.

According to the present specification there is provided the polymorphicForm E of Compound X, which has an X-ray powder diffraction pattern withat least two specific peaks at about 2-theta=17.9° and 14.8°.

According to the present specification there is provided the polymorphicForm E of Compound X, which has an X-ray powder diffraction pattern withspecific peaks at about 2-theta=17.9, 14.8, 20.9, 23.1, 16.2, 20.0,18.2, 12.3, 22.2 and 13.9°.

According to the present specification there is provided polymorphicForm E of Compound X which has an X-ray powder diffraction patternsubstantially the same as the X-ray powder diffraction pattern shown inFIG. 8.

According to the present specification there is provided polymorphicForm E of Compound X, which has an X-ray powder diffraction pattern withat least one specific peak at 2-theta=17.9° plus or minus 0.2° 2-theta.

According to the present specification there is provided a polymorphicForm E of Compound X, which has an X-ray powder diffraction pattern withat least one specific peak at 2-theta=14.8° plus or minus 0.2° 2-theta.

According to the present specification there is provided the polymorphicForm E of Compound X, which has an X-ray powder diffraction pattern withat least two specific peaks at 2-theta=17.9° and 14.8° wherein saidvalues may be plus or minus 0.2° 2-theta.

According to the present specification there is provided a polymorphicForm E of Compound X, which has an X-ray powder diffraction pattern withspecific peaks at 2-theta=17.9, 14.8, 20.9, 23.1, 16.2, 20.0, 18.2,12.3, 22.2 and 13.9° wherein said values may be plus or minus 0.2°2-theta.

It is to be understood that a suitable pharmaceutically acceptablepro-drug of a compound of the Formula (I) also forms an aspect of thepresent specification. Accordingly, the compounds of the specificationmay be administered in the form of a pro-drug, which is a compound thatis broken down in the human or animal body to release a compound of thespecification. A pro-drug may be used to alter the physical propertiesand/or the pharmacokinetic properties of a compound of thespecification. A pro-drug can be formed when the compound of thespecification contains a suitable group or substituent is to which aproperty-modifying group can be attached. Examples of pro-drugs includein-vivo cleavable ester or amide derivatives of the compound of theFormula (I).

Accordingly, one aspect of the present specification includes thosecompounds of Formula (I) as defined hereinbefore when made available byorganic synthesis and when made available within the human or animalbody by way of cleavage of a pro-drug thereof. Accordingly, the presentspecification includes those compounds of the Formula (I) that areproduced by organic synthetic means and also such compounds that areproduced in the human or animal body by way of metabolism of a precursorcompound, that is a compound of the Formula (I) may be asynthetically-produced compound or a metabolically-produced compound.

A suitable pharmaceutically acceptable pro-drug of a compound of theFormula (I) is one that is based on reasonable medical judgement asbeing suitable for administration to the human or animal body withoutundesirable pharmacological activities and without undue toxicity.

Various forms of pro-drug have been described, for example in thefollowing documents:—

-   -   a) Methods in Enzymology, Vol. 42, p. 309-396, edited by K.        Widder, et al. (Academic Press, 1985);    -   b) Design of Pro-drugs, edited by H. Bundgaard, (Elsevier,        1985);    -   c) A Textbook of Drug Design and Development, edited by        Krogsgaard-Larsen and H. Bundgaard, Chapter 5 “Design and        Application of Pro-drugs”, by H. Bundgaard p. 113-191 (1991);    -   d) H. Bundgaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992);    -   e) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77,        285 (1988);    -   f) N. Kakeya, et al., Chem. Pharm. Bull., 32, 692 (1984);    -   g) T. Higuchi and V. Stella, “Pro-Drugs as Novel Delivery        Systems”, A.C.S. Symposium Series, Volume 14; and    -   h) E. Roche (editor), “Bioreversible Carriers in Drug Design”,        Pergamon Press, 1987.

The in-vivo effects of a compound of the Formula (I) may be exerted inpart by one or more metabolites that are formed within the human oranimal body after administration of a compound of the Formula (I). Asstated hereinbefore, the in-vivo effects of a compound of the Formula(I) may also be exerted by way of metabolism of a precursor compound (apro-drug).

For the avoidance of doubt it is to be understood that where in thisspecification a group is qualified by ‘hereinbefore defined’ or ‘definedherein’ the said group encompasses the first occurring and broadestdefinition as well as each and all of the alternative definitions forthat group.

Another aspect of the present specification provides a process forpreparing a compound of the Formula (I), or a pharmaceuticallyacceptable salt thereof. A suitable process is illustrated by thefollowing representative process variants in which, unless otherwisestated, A, D, E, G, Q and R¹ to R¹² have any of the meanings definedhereinbefore. Necessary starting materials may be obtained by standardprocedures of organic chemistry. The preparation of such startingmaterials is described in conjunction with the following representativeprocess variants and within the accompanying Examples. Alternatively,necessary starting materials are obtainable by analogous procedures tothose illustrated which are within the ordinary skill of an organicchemist.

Compounds of Formula (I) where R¹³ is H may be made by, for example:

a) etherification of a suitable aryl or heteroaryl compound of Formula(II), where L is for example a halogen (such as iodine), or atrifluoromethanesulfonyl (triflate) group, or a boronic acid or ester,with an alcohol of Formula (III) using a suitable metal catalyst (forexample RockPhos 3rd Generation Precatalyst) in a suitable solvent (suchas toluene or DME) in the presence of a suitable base (such as cesiumcarbonate) and a suitable temperature (such as 90-120° C.); removal ofthe protecting group (PG) in Formula (II), such as THP, using acidconditions (such as anhydrous HCl in 1,4-dioxane) at suitabletemperature (such as 10-30° C.).

b) amination of a suitable aryl or heteroaryl compound of Formula (II),where L is for example a halogen (such as iodine), or atrifluoromethylsulfonyloxy (triflate) group, with an amine of Formula(IV) using a suitable metal catalyst (for example BrettPhos or RuPhos,and Pd₂(dba)₃) in a suitable solvent (for example 1,4-dioxane) in thepresence of a suitable base (for example cesium carbonate, sodiumtert-butoxide, or LiHMDS) at a suitable temperature (such as 90-130°C.); removal of the protecting group (PG), such as THP, using acidconditions (such as anhydrous HCl in 1,4-dioxane) at suitabletemperature (such as 10-30° C.).

c) alkylation of a suitable phenol or hydroxyl heteroaryl compound ofFormula (V) with an alcohol of Formula (III) via Mitsunobu reactionusing appropriate reagents (such as triphenylphosphine and diisopropyl(E)-diazene-1,2-dicarboxylate) in a suitable solvent (such as THF);removal of the protecting group (PG), such as THP, in Formula (VII),using acid conditions (such as anhydrous HCl in 1,4-dioxane) at suitabletemperature (such as 10-30° C.).

d) alkylation of a suitable aniline or heteroaryl amine or phenol orhydroxyl heteroaryl compound of Formula (VI) with a compound of Formula(VII) where LG is a leaving group (such as halide or mesylate), usingmild bases (for example DIPEA) in a suitable solvent (such as DMF orMeCN); removal of the protecting group (PG), such as THP, using acidconditions (such as anhydrous HCl in 1,4-dioxane) at suitabletemperature (such as 10-30° C.).

e) Alkylation of amines of Formula (VIII) with a suitable alkylatinggroup of Formula (IX) (wherein LG can be halide, as bromide, iodide orchloride, or may be some other suitable leaving group, such as mesylate)in a suitable solvent (such as DMF) in the presence of a suitable base(such as DIPEA) at a suitable temperature (such as 10-30° C.).

Compounds of formula (II) where R⁶ is not equal to hydrogen may be made,for example, from compounds of formula (X) by oxidation with a suitablereagent (for example bis(trifluoracetoxy)-iodobenzene) and treatmentwith an organometallic reagent (for example methyl magnesium bromidewhen R⁶ is methyl) in a suitable solvent (for example THF) at lowtemperature (typically −80 to −60° C.).

Compounds of formula (X) may be prepared by, for example, reaction of ananiline of Formula (XI) with suitable reagents to effect theconstruction of an indazole such as inorganic nitrite (such as sodiumnitrite) in organic acid (such as propionic acid) at low temperature(typically −20 to 0° C.) or alternatively an acid anhydride (such asacetic anhydride) in the presence of a suitable base (such as potassiumacetate) together with organic nitrite (such as isopentyl nitrite)optionally in the presence of a crown ether (such as 18-crown-6) in asuitable solvent (such as chloroform) at a suitable temperature (such as70° C.).

Compounds of formula (XI) may be made by reaction of a compound offormula (XII) with a compound of formula (XIII) under conditions knownin the art as suitable for Pictet-Spengler reactions, such as in thepresence of acid (such as acetic acid) and in a suitable solvent (forexample toluene or water) and a suitable temperature (such as 60-100°C.).

Compounds of formula (XII) may be prepared by functional groupinterconversions known to the art, for example aminations of halides offormula (XIV) from aryl halides (such as bromide) using a protectedamine (such as diphenylmethanimine) in the presence of a suitablecatalyst and ligand (such as bis(dibenzylideneacetone)palladium(0) andrac-2,2′-bis(diphenylphosphino)-1,1′-binaphthyl) in the presence of asuitable base (such as sodium tert-butoxide) in a suitable solvent (suchas toluene) at a suitable temperature (such as 80-100° C.).

Compounds of Formula (XIV) may be prepared by:

a) reaction of a compound of formula (XV) with an aldehyde of formula(XVI), in a suitable solvent (for example THF) in the presence of asuitable reducing agent (such as sodium triacetoxyborohydride) and at asuitable temperature (such as 20-30° C.);b) (i) reaction of a compound of formula (XV) with an acid of formula(XVII) under standard amide bond forming conditions (for example in thepresence of an amide coupling reagent (such as HATU) and a suitable base(such as triethylamine) in a suitable solvent (such as DMF)), followedby (ii) reduction of the resultant amide bond using a suitable reducingagent (such as borane) in a suitable solvent (such as THF) at a suitabletemperature (such as 60-70° C.);c) reaction of a compound of formula (XV) with a compound of formula(XVIII), wherein LG is a suitable leaving group (for example a halogenatom (such as bromo or chloro) or triflate), in the presence of asuitable base (such as diisopropylethylamine) in a suitable solvent (forexample DCM or dioxane) and at a suitable temperature (such as 20-85°C.).

Compounds of formula (XV) may be prepared by a number of methods knownto the art for the synthesis of chiral amines notably;

a) Ring opening of sulfamidates of Formula (XIX) according to the schemeshown below.

Step 1: Alkylation, e.g. n-butyllithium/THF/−78° C. to 0° C.Step 2: Removal of protection groups, e.g. anhydrous HCl in MeOH/DCM,rt.b) Phase transfer alklyation in the presence of a chiral catalyst (suchas(1S,2S,4S,5R)-2-((R)-(allyloxy)(quinolin-4-yl)methyl)-1-(anthracen-9-ylmethyl)-5-vinylquinuclidin-1-iumbromide) followed by functional group manipulation.

Step 1: Alkylation, e.g. Chiral catalyst, toluene/KOH, 0° C.Step 2: Functional group interconversion.

Compounds of Formula (XII) may be directly prepared by:

a) reaction of a compound of formula (XX) with an aldehyde of formula(XVI), in a suitable solvent (for example THF) in the presence of asuitable reducing agent (such as sodium triacetoxyborohydride) and at asuitable temperature (such as 20-30° C.);b) (i) reaction of a compound of formula (XX) with an acid of formula(XVII) under standard amide bond forming conditions (for example in thepresence of an amide coupling reagent (such as HATU) and a suitable base(such as triethylamine) in a suitable solvent (such as DMF)), followedby (ii) reduction of the resultant amide bond using a suitable reducingagent (such as borane) in a suitable solvent (such as THF) at a suitabletemperature (such as 60-70° C.);c) reaction of a compound of formula (XX) with a compound of formula(XIII), wherein LG is a suitable leaving group (for example a halogenatom (such as bromo or chloro) or triflate), in the presence of asuitable base (such as diisopropylethylamine) in a suitable solvent (forexample DCM or dioxane) and at a suitable temperature (such as 20-85°C.).

Compounds of Formula (XX) may be prepared through a reaction sequencestarting from a protected 3-bromo-2-methyl-aniline as shown below.

Step 1: Alkylation, e.g. n-butyllithium/THF/−78° C. to rt.Step 2: Removal of amine protection groups, e.g. anhydrous HCl inMeOH/DCM, rt.Step 3: Removal of aniline protection groups, e.g. refluxing inhydroxylamine.

Compounds of Formula (V) may be prepared through a sequence involvingPictet-Spengler cyclisation of a boronate ester-containing compound ofFormula (XIII) as described above to give a compound of Formula (XXI). Acompound of Formula (XXI) can be oxidized to a compound of Formula (V)using a suitable oxidant (such as hydrogen peroxide) in the presence ofa suitable base (such as sodium hydroxide) in a suitable solvent (suchas THF).

Compounds of Formula (VI, Q is NH) may be prepared through a sequenceinvolving Pictet-Spengler cyclisation of a nitro containing compound ofFormula (XIII) as described above to give a compound of Formula (XXII).A compound of Formula (XXII) can be reduced to a compound of Formula(VI) using suitable nitro reduction conditions (such as hydrogenation)in the presence of a suitable catalyst (such as platinum dioxide) in asuitable solvent (such as methanol).

Compounds of Formula (VIII), where Q is O, may be prepared from arylhalides of Formula (II) and tert-butyl 3-hydroxyazetidine-1-carboxylateusing a suitable metal catalyst (such as RockPhos 3rd GenerationPrecatalyst) in a suitable solvent (such as toluene or DME) in thepresence of a suitable base (such as cesium carbonate) at a suitabletemperature (such as 90-120° C.); the Boc protecting group may besubsequently removed under using an acid (such as trifluoroacetic acid)in a suitable solvent (such as DCM). Compounds of Formula (VIII) (Q isO) may also be prepared from a compound of Formula (V) under conditionsknown in the art as suitable for Mitsunobu reactions using appropriatereagents (such as triphenylphosphine and diisopropyl(E)-diazene-1,2-dicarboxylate) with tert-butyl3-hydroxyazetidine-1-carboxylate in a suitable solvent (such as THF).

Compounds of Formula (VIII), where Q is NH, may be prepared from arylhalides of Formula (II) and tert-butyl 3-aminoazetidine-1-carboxylateusing a suitable metal catalyst (for example for example RuPhos orBrettPhos and Pd₂(dba)₃) in a suitable solvent (for example 1,4-dioxane)in the presence of a suitable base (for example cesium carbonate, sodiumtert-butoxide, or LiHMDS) at a suitable temperature (such as 90-130°C.); the Boc protecting group may be subsequently removed using an acid(such as trifluoroacetic acid) in a suitable solvent (such as DCM).

Compounds of Formula (III) may be prepared by:

-   a) Alkylation reaction between 3-hydroxyazetidine and compounds of    Formula (IX) where LG for example a halogen or other leaving group    (such as mesyl group) in the presence of a suitable base, such as    cesium carbonate, in a suitable solvent, such as acetonitrile, at a    suitable temperature, such as 120° C., and in a suitable container,    such as a sealed tube.-   b) Reductive amination reaction between 3-hydroxyazetidine and    aldehyde or ketone compounds of Formula (XXIII) in the presence of a    suitable reducing reagent, such as sodium triacetoxyborohydride, in    a suitable solvent, such as DCM, in a suitable temperature, such as    10-30° C.

Compounds of Formula (IV) may be prepared by:

a) (i) Alkylation reaction between compound of Formula (XXIV), where PGis a pretecting group for example Boc, and compounds of Formula (IX)where LG is for example a halogen or other leaving group, such asmesylate, in the presence of a suitable base, such as DIPEA, in asuitable solvent, such as 1,4-dioxane, at a suitable temperature, suchas 10-30° C. (ii) Removal of the protection group under suitableconditions, such as acidic conditions for the removal of Boc.

b) (i) Reductive amination reaction between compounds of Formula (XXIV)and aldehyde or ketone compounds of Formula (XXIII) in the presence of asuitable reducing reagent, such as sodium triacetoxyborohydride, in asuitable solvent, such as DCM, in a suitable temperature, such as 10-30°C. (ii) Removal of the protection group under suitable conditions, suchas acidic conditions for the removal of Boc.

It is to be understood that other permutations of the process steps inthe process variants described above are also possible.

It will also be appreciated that, in some of the reactions mentionedhereinbefore, it may be necessary or desirable to protect any sensitivefunctionalities in the compounds. The instances where protection isnecessary or desirable, and suitable methods for protection, are knownto those skilled in the art. Conventional protecting groups may be usedin accordance with standard practice (for illustration see T. W. Green,Protective Groups in Organic Synthesis, John Wiley and Sons, 1991).Thus, if reactants include groups such as amino, carboxy or hydroxy, itmay be desirable to protect the group in some of the reactions mentionedherein.

A suitable protecting group for an amino or alkylamino group is, forexample, an acyl group, for example an alkanoyl group such as acetyl, analkoxycarbonyl group, for example a methoxycarbonyl, ethoxycarbonyl ort-butoxycarbonyl group, an arylmethoxycarbonyl group, for examplebenzyloxycarbonyl, or an aroyl group, for example benzoyl. Thedeprotection conditions for the above protecting groups necessarily varywith the choice of protecting group. Thus, for example, an acyl groupsuch as an alkanoyl or alkoxycarbonyl group or an aroyl group may beremoved for example, by hydrolysis with a suitable base such as analkali metal hydroxide, for example lithium or sodium hydroxide.Alternatively an alkoxycarbonyl group such as a t-butoxycarbonyl groupmay be removed, for example, by treatment with a suitable acid ashydrochloric, sulphuric, formic, phosphoric or trifluoroacetic acid, andan arylmethoxycarbonyl group such as a benzyloxycarbonyl group may beremoved, for example, by hydrogenation over a catalyst such aspalladium-on-carbon, or by treatment with a Lewis acid, such as borontris(trifluoroacetate). A suitable alternative protecting group for aprimary amino group is, for example, a phthaloyl group, which may beremoved by treatment with an alkylamine, for exampledimethylaminopropylamine, or hydrazine.

A suitable protecting group for a hydroxy group is, for example, an acylgroup, for example an alkanoyl group such as acetyl, an aroyl group, forexample benzoyl, an arylmethyl group, for example benzyl, or a trialkylor diarylalkyl silane, such as TBDMS or TBDPS. The deprotectionconditions for the above protecting groups will necessarily vary withthe choice of protecting group. Thus, for example, an acyl group such asan alkanoyl or an aroyl group may be removed, for example, by hydrolysiswith a suitable base such as an alkali metal hydroxide, for examplelithium or sodium hydroxide. Alternatively an arylmethyl group such as abenzyl group may be removed, for example, by hydrogenation over acatalyst such as palladium-on-carbon.

A suitable protecting group for a carboxy group is, for example, anesterifying group, for example a methyl or an ethyl group which may beremoved, for example, by hydrolysis with a base such as sodiumhydroxide, or for example a t-butyl group which may be removed, forexample, by treatment with an acid, such as trifluoroacetic acid, or forexample a benzyl group which may be removed, for example, byhydrogenation over a catalyst such as palladium-on-carbon.

The protecting groups may be removed at any convenient stage in thesynthesis using conventional techniques well known in the chemical art.

Certain of the intermediates defined herein are novel and these areprovided as further features of the specification.

In one embodiment there is provided a compound of Formula (XXV), or asalt thereof:

wherein L is bromo, chloro, iodo or trifluoromethanesulfonyl.

In a further embodiment L is bromo.

Biological Assays

The following assays were used to measure the effects of the compoundsof the present specification.

ERα Binding Assay

The ability of compounds to bind to isolated Estrogen Receptor AlphaLigand binding domain (ER alpha-LBD (GST)) was assessed in competitionassays using a LanthaScreen™ Time-Resolved Fluorescence Resonance EnergyTransfer (TR-FRET) detection end-point. For the LanthaScreen TR-FRETendpoint, a suitable fluorophore (Fluormone ES2, ThermoFisher, Productcode P2645) and recombinant human Estrogen Receptor alpha ligand bindingdomain, residues 307-554 (expressed and purified in-house) were used tomeasure compound binding. The assay principle is that ER alpha-LBD (GST)is added to a fluorescent ligand to form a receptor/fluorophore complex.A terbium-labelled anti-GST antibody (Product code PV3551) is used toindirectly label the receptor by binding to its GST tag, and competitivebinding is detected by a test compound's ability to displace thefluorescent ligand, resulting in a loss of TR-FRET signal between theTb-anti-GST antibody and the tracer. The assay was performed as followswith all reagent additions carried out using the Beckman CoulterBioRAPTR FRD microfluidic workstation:

-   -   1. Acoustic dispense 120 nL of the test compound into a black        low volume 384 well assay plates.    -   2. Prepare 1×ER alpha-LBD/Tb-antiGST Ab in ES2 screening buffer        and incubate for 15 minutes.    -   3. Dispense 6 μL of the 1×AR-LBD/Tb-anti-GST Ab reagent into        each well of the assay plate followed by 6 μL of Fluorophore        reagent into each well of the assay plate    -   4. Cover the assay plate to protect the reagents from light and        evaporation, and incubate at room temperature for 4 hours.    -   5. Excite at 337 nm and measure the fluorescent emission signal        of each well at 490 nm and 520 nm using the BMG PheraSTAR.

Compounds were dosed directly from a compound source microplatecontaining serially diluted compound (4 wells containing 10 mM, 0.1 mM,1 mM and 10 nM final compound respectively) to an assay microplate usingthe Labcyte Echo 550. The Echo 550 is a liquid handler that usesacoustic technology to perform direct microplate-to-microplate transfersof DMSO compound solutions and the system can be programmed to transfermultiple small nL volumes of compound from the different source platewells to give the desired serial dilution of compound in the assay whichis then back-filled to normalise the DMSO concentration across thedilution range.

In total 120 nL of compound plus DMSO were added to each well andcompounds were tested in a 12-point concentration response format over afinal compound concentration range of 10, 2.917, 1.042, 0.2083, 0.1,0.0292, 0.0104, 0.002083, 0.001, 0.0002917, 0.0001042, and 0.00001 μMrespectively. TR-FRET dose response data obtained with each compound wasexported into a suitable software package (such as Origin or Genedata)to perform curve fitting analysis. Competitive ER alpha binding wasexpressed as an IC₅₀ value. This was determined by calculation of theconcentration of compound that was required to give a 50% reduction intracer compound binding to ER alpha-LBD.

MCF-7 ER Down-Regulation Assay

The ability of compounds to down-regulate Estrogen Receptor (ER) numberswas assessed in a cell based immuno-fluorescence assay using the MCF-7human ductal carcinoma breast cell line. MCF-7 cells were reviveddirectly from a cryovial (approx 5×10⁶ cells) in Assay Medium (phenolred free Dulbecco's Modified Eagle's medium (DMEM); Sigma D5921)containing 2 mM L-Glutamine and 5% (v/v) Charcoal/Dextran treated foetalcalf serum. Cells were syringed once using a sterile 18 G×1.5 inch(1.2×40 mm) broad gauge needle and cell density was measured using aCoulter Counter (Beckman) Cells were further diluted in Assay Medium toa density of 3.75×10⁴ cells per mL and 40 μL per well added totransparent bottomed, black, tissue culture-treated 384 well plates(Costar, No. 3712) using a Thermo Scientific Matrix WellMate or ThermoMultidrop. Following cell seeding, plates were incubated overnight at37° C., 5% CO₂ (Liconic carousel incubator). Test data was generatedusing the LabCyte Echo™ model 555 compound reformatter which is part ofan automated workcell (Integrated Echo 2 workcell). Compound stocksolutions (10 mM) of the test compounds were used to generate a 384 wellcompound dosing plate (Labcyte P-05525-CV1). 40 μL of each of the 10 mMcompound stock solutions was dispensed into the first quadrant well andthen 1:100 step-wise serial dilutions in DMSO were performed using aHydra II (MATRIX UK) liquid handling unit to give 40 μL of dilutedcompound into quadrant wells 2 (0.1 mM), 3 (1 μM) and 4 (0.01 μM),respectively. 40 μL of DMSO added to wells in row P on the source plateallowed for DMSO normalisation across the dose range. To dose thecontrol wells 40 μL of DMSO was added to row O1 and 40 μL of 100 μMfulvestrant in DMSO was added to row O3 on the compound source plate.

The Echo uses acoustic technology to perform directmicroplate-to-microplate transfers of DMSO compound solutions to assayplates. The system can be programmed to transfer volumes as low as 2.5nL in multiple increments between microplates and in so doing generatesa serial dilution of compound in the assay plate which is thenback-filled to normalise the DMSO concentration across the dilutionrange. Compounds were dispensed onto the cell plates with a compoundsource plate prepared as above producing a 12 point duplicate 3 μM to 3μM dose range with 3-fold dilutions and one final 10-fold dilution usingthe Integrated Echo 2 workcell. The maximum signal control wells weredosed with DMSO to give a final concentration of 0.3%, and the minimumsignal control wells were dosed with fulvestrant to give a finalconcentration of 100 nM accordingly. Plates were further incubated for18-22 hours at 37° C., 5% CO₂ and then fixed by the addition of 20 μL of11.1% (v/v) formaldehyde solution (in phosphate buffered saline (PBS))giving a final formaldehyde concentration of 3.7% (v/v). Cells werefixed at room temperature for 20 mins before being washed two times with250 μL PBS/Proclin (PBS with a Biocide preservative) using a BioTekplatewasher, 40 μL of PBS/Proclin was then added to all wells and theplates stored at 4° C. The fixing method described above was carried outon the Integrated Echo 2 workcell Immunostaining was performed using anautomated AutoElisa workcell. The PBS/Proclin was aspirated from allwells and the cells permeabilised with 40 μL PBS containing 0.5% Tween™20 (v/v) for 1 hour at room temperature. The plates were washed threetimes in 250 μL of PBS/0.05% (v/v) Tween 20 with Proclin (PBST with aBiocide preservative) and then 20 μL of ERα (SP1) Rabbit monoclonalantibody (Thermofisher) 1:1000 in PBS/Tween™/3% (w/v) Bovine SerumAlbumin was added. The plates were incubated overnight at 4° C. (Liconiccarousel incubator) and then washed three times in 250 μL of PBS/0.05%(v/v) Tween™ 20 with Proclin (PBST). The plates were then incubated with20 μL/well of a goat anti-rabbit IgG AlexaFluor 594 or goat anti-rabbitAlexaFluor 488 antibody (Molecular Probes) with Hoechst at 1:5000 inPBS/Tween™/3% (w/v) Bovine Serum Albumin for 1 hour at room temperature.The plates were then washed three times in 250 μL of PBS/0.05% (v/v)Tween™ 20 with Proclin (PB ST with a Biocide preservative). 20 μL of PBSwas added to each well and the plates covered with a black plate sealand stored at 4° C. before being read. Plates were read using aCellomics Arrayscan reading the 594 nm (24 hr time point) or 488 nm (5hr timepoint) fluorescence to measure the ERα receptor level in eachwell. The mean total intensity was normalized for cell number giving thetotal intensity per cell. The data was exported into a suitable softwarepackage (such as Origin) to perform curve fitting analysis.Down-regulation of the ERα receptor was expressed as an IC₅₀ value andwas determined by calculation of the concentration of compound that wasrequired to give a 50% reduction of the average maximum Total Intensitysignal.

The data shown in Table A were generated (the data below may be a resultfrom a single experiment or an average of two or more experiments):

TABLE A ER binding IC50 ER down regulation Example value (nM) IC50 value(nM)¹  1 3.7 0.10  2 2.8 3.1  3 6.5 0.64  4 6.9 0.34  5 2.2 0.077  6 8.80.35  7 8.4 0.4  8 2.0 0.055  9 360 >300 10 8 0.26 11 20 0.094 12 2.40.25 (83%) 13 3.5 0.11 14 6.6 >300 15 9.5 0.33 (84%) 16 6.9 0.54 17 2.40.16 18 29 0.42 19 2.6 0.063 20 15 0.36 21 2.1 0.098 22 5.7 0.062 23 150.15 24 10 0.21 25 4.1 0.33 26 6.7 0.52 27 0.81 0.05 28 5.7 0.065 29 1.60.2 30 1.8 0.28 31 13 0.54 32 2.1 0.096 33 1.4 0.1 34 1.9 0.22 35 1.50.17 36 1.8 0.12 37 7.2 4.1 38 0.69 0.19 39 0.94 0.048 40 1.3 0.1 41 2.40.17 42 3.8 0.28 43 4.1 0.25 44 9.6 0.91 45 1.6 0.3 46 3.6 0.4 47 1.20.41 48 1.5 0.078 49 1.5 0.075 50 2.7 0.47 51 2.8 0.68 52 11 0.21 53 5.70.26 54 160 >300 55 8.2 1.1 56 8.2 0.58 57 5 0.36 58 2.1 0.56 59 7.2 1.660 4.7 0.19 61 0.88 0.13 62 1 0.13 63 73 0.31 64 6.1 0.22 65 5.3 1 666.2 0.44 67 5 0.27 68 15 0.4 69 8.9 0.55 70 4.3 0.31 71 2 0.17 72 2.90.51 73 3.4 0.77 ¹Compounds which are active in the ER down-regulationassay show downregulation values > 90% in the assay unless otherwisestated, in which case the % downregulation is shown in parentheses.

Western Blotting Assay

The ability of compounds to down-regulate estrogen receptor (ER) wasassessed by western blotting using human breast cancer cell lines (MCF-7and CAMA-1). Cells were plated into 12-well tissue culture-treatedplates at 0.5×10⁶/well in phenol red-free RPMI containing 2 mML-glutamine and 5% (v/v) charcoal treated foetal calf serum (F6765,Sigma). Cells were incubated with compounds (100 nM) or vehicle control(0.1% DMSO) for 48 h at 37° C., 5% CO₂ before washing once with PBS andlysing with 80 μl lysis buffer (25 mM Tris/HCl, 3 mM EDTA, 3 mM EGTA, 50mM NaF, 2 mM sodium orthovanadate, 0.27 M sucrose, 10 mMβ-glycerophosphate, 5 mM sodium pyrophosphate, 0.5% TritonX-100, pH 6.8)on ice.

Cells were scraped, sonicated and centrifuged prior to performing aprotein assay (DC Bio-Rad Protein kit, 500-0116) and making samples to aprotein concentration of 1-2 mg/ml in lysis buffer containing 1×LDSSample Buffer (NP0007, Invitrogen) and 1× NuPAGE sample reducing agent(NP0009, Invitrogen). Samples were boiled for 10 min at 95° C. and thenfrozen at −20° C. until ready for use.

10-20 μg protein was loaded onto 26-well Criterion gels (BioRad345-0034). Gels were run at 125 V for 1 hr 25 min in running buffer (24mM Tris Base Sigma, 192 mM Glycine, 3.5 mM SDS, made up in distilledwater). Gels were then transferred at 30V for 2 hr in transfer buffer(25 mM Tris, 192 mM Glycine, 20% (v/v) methanol, pH 8.3, made up indistilled water) onto nitrocellulose membrane. The blot was stained withPonceau S (P7170, Sigma) and cut according to appropriate molecularweight markers.

Membranes were blocked for 1 hour at room temp in 5% Marvel (w/v) inphosphate-buffered saline containing 0.05% Tween™ 20 (PBS/Tween). Blotswere then incubated with anti-ERα (SP1) rabbit monoclonal antibody(Thermofisher) diluted 1:1000 at 4° C. overnight (with gentle shaking)followed by several washes with PBS/Tween. Secondary anti-rabbit HRPantibody (7074, CST) diluted 1:2000 dilution was incubated for 2 h atroom temperature (with gentle shaking) followed by several washes withPBS/Tween. All antibodies were made up in 5% Marvel (w/v) in PBS/Tween.

The immunoblots were developed using Pierce WestDura chemiluminescentreagents (Thermo Scientific 34076) and developed/quantified on the G-boxusing Syngene software. Down-regulation of the ERα receptor wasnormalised to the vehicle control (0% down-regulation) and the 100 nMfulvestrant control (100% down-regulation) run within the same gel.

Table B shows the data generated for selected Examples (the data belowmay be a result from a single experiment or an average of two or moreexperiments):

TABLE B CAMA1 Western MCF7 Western Example % ER deg vs Fv % ER deg vs Fv 1 92 91  2 95 93  3 102 90  4 61 70  5 109 99  6 84 89  7 90 90  8 100101  9 25 −13 16 105 104 17 96 92 18 95 103 19 94 92 21 101 99 25 96 9826 97 97 27 108 110 28 110 95 29 106 93 30 98 96 31 95 103 32 89 89 33102 98 34 82 85 40 120 98 42 79 92 46 102 105 48 94 93 49 99 98 53 46 8156 52 61 59 102 104 64 89 86 69 98 95 70 100 102

Human Hepatocyte Assay

The metabolic stability of compounds in human hepatocytes was assessedusing the following protocol:

-   -   1. Prepare 10 mM stock solutions of compound and control        compounds in appropriate solvent (DMSO). Place incubation medium        (L-15Medium) in a 37° C. water bath, and allow warming for at        least 15 minutes prior to use.    -   2. Add 80 μL of acetonitrile to each well of the 96-well deep        well plate (quenching plate).    -   3. In a new 96-well plate, dilute the 10 mM test compounds and        the control compounds to 100 μM by combining 198 μL of        acetonitrile and 2 μL of 10 mM stock.    -   4. Remove a vial of cryopreserved (less than −150° C.) human        hepatocytes (LiverPool™ 10 Donor Human hepatocytes obtained from        Celsis IVT. Chicago, Ill. (Product No. S01205)) from storage,        ensuring that vials remain at cryogenic temperatures until        thawing process ensues. As quickly as possible, thaw the cells        by placing the vial in a 37° C. water bath and gently shaking        the vials. Vials should remain in water bath until all ice        crystals have dissolved and are no longer visible. After thawing        is complete, spray vial with 70% ethanol, transfer the vial to a        bio-safety cabinet.    -   5. Open the vial and pour the contents into the 50 mL conical        tube containing thawing medium. Place the 50 mL conical tube        into a centrifuge and spin at 100 g for 10 minutes. Upon        completion of spin, aspirate thawing medium and resuspend        hepatocytes in enough incubation medium to yield ˜1.5×10⁶        cells/mL.    -   6. Using Cellometer® Vision, count cells and determine the        viable cell density. Cells with poor viability (<80% viability)        are not acceptable for use. Dilute cells with incubation medium        to a working cell density of 1.0×10⁶ viable cells/mL.    -   7. Transfer 247.5 μL of hepatocytes into each well of a 96-well        cell culture plate. Place the plate on Eppendorf Thermomixer        Comfort plate shaker to allow the hepatocytes to warm for 10        minutes.    -   8. Add 2.5 μL of 100 μM test compound or control compounds into        an incubation well containing cells, mix to achieve a homogenous        suspension at 0.5 min, which when achieved, will define the 0.5        min time point. At the 0.5 min time, transfer 20 μL of incubated        mixture to wells in a “Quenching plate” followed by vortexing.    -   9. Incubate the plate at 37° C. at 900 rpm on an Eppendorf        Thermomixer Comfort plate shaker. At 5, 15, 30, 45, 60, 80, 100        and 120 min, mix the incubation system and transfer samples of        20 μL incubated mixture at each time point to wells in a        separate “Quenching plate” followed by vortexing.    -   10. Centrifuge the quenching plates for 20 minutes at 4,000 rpm.        4 different compounds are pooled into one cassette and used for        LC/MS/MS analysis.

All calculations were carried out using Microsoft Excel. Peak areas weredetermined from extracted ion chromatograms. In vitro intrinsicclearance (in vitro Cl_(int), in L/min/10⁶ cells) of parent compound wasdetermined by regression analysis of the Ln percent parent disappearancevs. time curve. The in vitro intrinsic clearance (in vitro Cl_(int), inL/min/10⁶ cells) was determined from the slope value using the followingequation and is shown in Table C for selected examples:

in vitro Cl_(int)=kV/N

V=incubation volume (0.25 mL);

N=number of hepatocytes per well (0.25×10⁶ cells).

TABLE C Cl_(int) (μL/min/10⁶ Example cells)  1 11  2 6  3 5  6 2 11 7 176 18 3 19 5 21 6 25 5 28 5 48 4

Physical Properties log D

The lipophilicity of a drug is an important physical property which mayinfluence many biological and metabolic properties of a compound, forexample the absorption, distribution, metabolism, excretion and toxicityprofiles of a compound. The distribution coefficient between 1-octanoland aqueous buffer, Log DO/W, at pH 7.4, is the most commonly usedmeasurement of the lipophilicity of a compound. The current method formeasurement of Log DO/W is based on the traditional shake flasktechnique, but with the modification of measuring compounds in mixturesof ten at a time using UPLC with quantitative mass spectrometry (MS) asa method to measure the relative octanol and aqueous concentrations. Themaximum capacity is 379 project compounds (48 pools with 10 compoundsincl. three QC compounds) per experiment. 2 quality control (QC)samples, Cyclobenzaprine with moderate Log D and Nicardipine high Log Dis used in all pools to ensure good quality. An additional QC sampleCaffeine, with low Log D, are used and randomly placed in all runs. Themethod has been thoroughly validated against the previous shake flaskmethodologies.

Solubility

In order for an oral compound to reach the site of action, and in orderfor oral absorption from the gut to occur, that compound must be insolution, and therefore compounds which possess high intrinsicsolubility may be more suitable for pharmaceutical use. Thethermodynamic solubility of a research compound is measured understandard conditions. It is a shake-flask approach that uses 10 mM DMSOsolutions which are supplied from the Compound Managements liquid storeand is a high throughput method. The dried compounds are equilibrated inan aqueous phosphate buffer (pH 7.4) for 24 hours at 25° C., the portionwith the dissolved compound is then separated from the remains. Thesolutions are analyzed and quantified using UPLC/MS/MS, QC-samples areincorporated in each assay-run to ensure the quality of the assay.

Human Plasma Protein Binding

Hunan plasma protein binding is a key factor in controlling the amountof free (unbound) drug available for binding to target and hence playsan important role in the observed efficacy of drugs in vivo. Therefore,compounds which possess high free fraction (low levels of plasma proteinbinding) may exhibit enhanced efficacy relative to a compound withsimilar potency and exposure levels. The automated equilibrium dialysisassay in human plasma uses the RED (Rapid Equilibrium Dialysis) Deviceand sample handling. The assay generally runs over two to three daysincluding delivery of results. After dialysis for 18 hours, plasma andbuffer samples are prepared for analysis by liquid chromatography andmass spectrometry. Samples are generally tested in singlicates andquantified by LC/MSMS by using a 7-point calibration curve in plasma.The compounds are pooled together in plasma pools up to 10 compounds.Three reference compounds are used in each run, Propranolol, Metoprololand Warfarin. Warfarin is used as a control in each pool and Propranololand Metoprolol are placed randomly in each run. An in-house Excel macrois used for preparation of files for the robot and the mass spectrometerand is also used for the calculations of fraction unbound (fu %) inplasma.

Table D shows the data for log D, solubility and plasma protein bindinggenerated for selected Examples (the data below may be a result from asingle experiment or an average of two or more experiments):

TABLE D Human plasma LogD protein binding Solubility Example pH 7.4 (%free) (μM)  1 3.4 8.3 379  2 2.6 28 217  3 2.5 26 >941  4 2.7 21 991  53.3 8.6 297  6 2.4 34 >947  7 3.5 11 470  8 4.4 0.99 90  9 1 21 >1000 102.8 23 634 11 3.9 2 186 12 3.9 2.7 77 13 4 6.4 78 14 2.6 16 897 15 2.534 821 16 2 64 982 17 2.9 23 833 18 2.6 34 910 19 3.7 4.3 152 20 2.8 29730 21 3.1 12 634 22 4 4.6 70 23 3.1 9.5 339 25 2.5 40 964 26 3 22 25827 3.8 3.8 229 28 3.6 6 150 29 3 16 779 30 3 16 754 31 2 44 >942 32 3.56.5 692 33 3.6 5.4 466 34 3.2 15 471 37 2.6 25 961 38 3.1 13 794 39 3.76.8 257 40 3.5 6.4 479 41 4.1 2.2 120 42 2.4 NT 712 44 3.2 10 998 45 3.96.2 172 46 2.5 31 >1000 47 2.6 NT >1000 48 3.1 10 582 49 3.8 NT 163 502.1 NT 748 51 2.7 28 >1000 52 3 19 864 53 3.2 11 691 54 1.4 57 >1000 552.1 NT 987 56 3.6 NT 377 57 2.9 NT 615 58 2.4 NT 633 59 2.4 33 971 602.5 23 593 61 2.7 29 848 62 2.8 19 784 63 3.2 10 870 64 3.2 14 474 652.4 NT 790 66 2.3 NT 931 67 2.7 19 914 68 2.4 37 561 69 2.9 37 >1000 702.9 24 857 71 3.2 NT 666 72 3.3 14 738 73 3 22 925 NT = not testedhERG Binding Assay

hERG (human ether go go-related gene) potassium channels are essentialfor normal electrical activity in the heart. Arrhythmia can be inducedby a blockage of hERG channels by a diverse group of drugs. This sideeffect is a common reason for drug failure in preclinical safety trials[Sanguinetti et al., Nature., 2006, 440, 463-469.] and thereforeminimisation of hERG channel blocking activity may be a desirableproperty for drug candidates.

The purpose of the hERG binding assay is to evaluate the effects of testcompounds on the voltage-dependent potassium channel encoded by thehuman ether go go-related gene (hERG) using a constitutively expressingCHO cell line on the Nanion Syncropatch 384PE automated patch clampsystem.

The assay was conducted as follows with all reagents used at roomtemperature unless otherwise stated.

Reagent Preparations Include:

1. Internal “IC700” solution used to perfuse the underside of chip (inmM), KF 130, KCl 20, MgCl2 1, EGTA 10 and HEPES 10, (all Sigma-Aldrich;pH 7.2-7.3 using 10 M KOH, 320 mOsm) and supplemented with 25 □M escin.2. External and cell buffer (in mM), NaCl 137, KCl 4, HEPES 10,D-glucose 10, CaCl2 2, MgCl2 1 (pH7.4, NaOH)3. NMDG “reference” buffer used to establish a stable baseline prior tothe addition of test compounds, NaCl 80, KCl 4, CaCl2 2, MgCl2 1, NMDGCl 60, D-Glucose monohydrate 5, HEPES 10 (pH7.4 NaOH 298 mOsm)4. Seal enhancer used to improve seal quality of cells, NaCl 80, KCl 3,CaCl2 10, HEPES 10, MgCl2 1 (pH7.4 NaOH)

Cell Preparations:

1. If using cell culture; cells to be incubated at 30° C. forapproximately 4-6 days prior to being used. Day of assay lift cellsusing accutase and re-suspend in 20 ml cell buffer to a density of 0.8to 1e6 cells/ml.2. If using assay ready cryovials; rapidly thaw two cryovials at 37° C.and slowly pipette into 23 ml external solution3. All cell preps to be incubated for 15 min on the shaking cell hotelset to 10° C. prior to starting assay

Compound Preparations:

All compounds were acoustically dispensed in quadruplicate using aLabcyte Echo. A 10 mM stock solution is used to generate 6 compoundsource plates each at a different concentration to allow cumulativedosing onto cells (0.03167 mM, followed by 0.1 mM, then 0.3167 mM, 1 mM,3.167 mM, 10 mM,). 90 μl of reference buffer is added to each well ofthe source plates containing 600 nl of compound for a final compoundconcentration of 0.1 μM, 0.39 μM, 1.2 μM, 3.9 μM, 12.5 μM and 39.6 μMrespectively. hERG assay (all dispense steps are performed using theliquid handling set up on the Nanion syncropatch)

1. Fill 384 well medium resistance 4 hole chips with 40 μl externalbuffer and perfuse internal buffer to the underside of plate.2. Dispense 20 μl of cells into each well of the chip followed by 20 μlof seal enhancer.3. Remove 40 μl of reagent from each well to the wash station, leaving aresidual volume of 40 μl4. Dispense 40 μl of reference buffer with a removal step of 40 μl after3 min, repeat this step.5. Dispense 40 μl of compound plate 1 (0.03167 mM), ‘real time’recordings for 3 min exposure prior to removal of 40 μl. This step isrepeated for 5 further subsequent compound plates in increasingconcentrations to generate a cumulative concentration-effect curve ineach well of the Syncropatch chip.

hERG-mediated currents were elicited using a voltage step protocolconsisting of acontinuous holding voltage of −80 mV, with a 500 ms stepto 60 mV followed by a 500 ms step to −40 mV every 15 seconds. hERGcurrent magnitude was measured automatically from the leak-subtractedtraces by the Nanion software by taking the peak of the hERG “tail”current at −40 mV every 15 seconds and taking the last three of theseresponses for each concentration to generate the concentration-effectcurve.

Calculation of results is performed using APC package within Genedata.For the routine normalization of well data with Neutral and Inhibitorcontrol well groups as reference, GeneData Assay Analyzer uses thefollowing equation to normalize the signal values to the desired signalrange:

${N(x)} = {{CR} + {\frac{{x -} < {cr} >}{< {sr} > {- {< {cr} >}}}( {{SR} - {CR}} )}}$

x is the measured raw signal value of a well<cr> is the median of the measured signal values for the CentralReference (Neutral) wells on a plate<sr> is the median of the measured signal values for the Scale Reference(Inhibitor) wells on a plateCR is the desired median normalized value for the Central Reference(Neutral)SR is the desired median normalized value for the Scale Reference(Inhibitor)

Table E shows the hERG binding data for selected Examples (the databelow may be a result from a single experiment or an average of two ormore experiments):

TABLE E hERG IC₅₀ Example (μM)  1 10  2 >40  3 >36  4 17  5 13  6 9.116 >40 17 22 18 >38 19 5.4 21 5.3 25 28 27 7.9 28 7.8 29 27 30 >3331 >40 32 14 33 14 34 7.7 37 20 38 18 39 14 40 4.5 41 6.5 42 7.9 45 4.246 >40 49 8.2 50 13 51 9.1 52 2 55 >40 57 16 58 >40 59 >40 60 11 61 >4062 >40 63 13 64 13 65 31 66 26 67 14 68 >40 69 24 70 24 73 12

Permeability

In order to maximize oral absorption, a drug must have sufficienttransmembrane flux as well as avoid efflux by P-glycoprotein. The mostwidely used system for predicting oral absorption is by determination ofthe permeation rate of compounds through monolayers of a human colonadenocarcinoma cell line Caco-2.

Human Caco-2 Bidirectional Permeability A to B and B to A

An automated assay was used to determine the bidirectional permeability(efflux and uptake) of compounds in Caco-2 cells carried out over 2hours at pH 7.4. Samples were analyzed through LC/MS/MS to estimate theapparent permeability coefficients (Papp) of compounds across Caco-2cell monolayers and results are quoted in units of ×10⁻⁶ cm/s. Theefflux ratio (ER) can be determined using the following equation:

ER=P _(app (B-A)) /P _(app (A-B))

Where P_(app (B-A)) indicates the apparent permeability coefficient inbasolateral to apical direction, and P_(app (A-B)) indicates theapparent permeability coefficient in apical to basolateral direction.

Human Caco-2 Passive Permeability A to B Papp

An automated assay was used to determine the passive permeability ofcompounds in Caco-2 cell monolayers carried out over 2 hours with anapical pH of 6.5 and basolateral pH of 7.4. The Caco-2 AB inhibitionassay is carried out with chemical inhibition of the three major effluxtransporters ABCB1 (P-gp), ABCG2 (BCRP) and ABCC2 (MRP2) in Caco-2cells. Incubation of both apical and basolateral is carried out with acocktail of inhibitors (50 μM quinidine, 20 μM sulfasalazine and 100 μMbenzbromarone). Samples were analyzed through LC/MS/MS to estimate theapparent permeability coefficients (Papp) of compounds across Caco-2cell monolayers and results are quoted in units of ×10⁻⁶ cm/s.

Table F shows the data for permeability generated for selected Examples(the data below may be a result from a single experiment or an averageof two or more experiments):

TABLE F Bidirectional Bidirectional Passive Caco-2 Papp Caco-2 Caco-2Papp Example (×10⁻⁶ cm/s) efflux ratio (×10⁻⁶ cm/s)  1 0.9 9.4 NT  2 3.98.4 NT  3 2.3 9.2 NT  6 0.9 28 1.8 11 1.6 1.4 NT 16 1.6 9.7 1.7 17 4.75.5 13.3 19 4.2 2.6 NT 21 4.0 1.6 14.1 25 5.0 7.4 5.9 27 5.4 0.9 NT 281.2 3.8 24 29 12 0.9 24 30 2.7 7.8 NT 31 0.9 9.4 NT 32 3.4 1.5 NT 33 2.61.5 NT 47 18 1.1 NT 60 0.7 34 NT 64 2.0 5.6 NT NT = not tested

Human Parental MCF7 Xenograft Anti-Tumour Efficacy in Mouse

To determine the effect of Example 17 on the growth of the MCF7xenografts, the following study was performed. MCF7 cells (ATCC) weregrown in vitro in exponential phase prior to implant. Briefly, male SCIDmice weighing 18 g or more (Envigo UK) were implanted subcutaneously onthe back with estrogen pellets (0.5 mg, 21 day release from InnovativeResearch of America) under recoverable anaesthetic. One day later micewere inoculated subcuaneously on the left flank with 5 million MCF7cells, prepared as 0.1 ml cell suspension in 1:1 RPMI (Gibco, LifeTechnologies) and matrigel (Corning). When tumours reached ˜250 mm³ micewere randomised into groups of 9 mice (12 mice for vehicle control) andstarted receiving drug treatment. Compounds were prepared in vehicle(40% Tetraethylene Glycol (v/v), 7.5% Captisol (w/v) in water forinjection) and given orally in a volume of 10 ml/kg once a day for 21days at 0.5 mg/kg to 50 mg/kg. Tumours were measured twice a week andtumour volume calculated using the elliptical formula(pi/6×width×width×length). Data represent the geomean of tumour volumerelative to the tumour volume on day of randomisation. Error bars arethe 95% confidence interval (Graphpad Prism). This study demonstratedthat doses of 10 mg/kg and above gave tumour regression (FIG. 12).

Human Y537S ESR1 Mutant MCF7 Xenograft Anti-Tumour Efficacy in Mouse

To determine the effect of Example 17 on the growth of the xenograftsderived from MCF7 cells genetically engineered to express Y537S ESR1,the following study was performed. Y537S ESR1 MCF7 cells were created bygenome editing and express only Y537S ESR1 (Ladd, et al., Oncotarget,2016, 7:54120-54136). Briefly, male SCID mice weighing 18 g or more(Envigo UK) were inoculated subcuaneously on the left flank with 5million Y537S ESR1 MCF7 cells, prepared as 0.1 ml cell suspension in 1:1RPMI (Gibco, Life Technologies) and matrigel (Corning). When tumoursreached ˜250 mm³ mice were randomised into groups of 9 mice (12 mice forvehicle control) and started receiving drug treatment. Compounds wereprepared in vehicle (40% Tetraethylene Glycol (v/v), 7.5% Captisol (w/v)in water for injection) and given orally in a volume of 10 ml/kg once aday for 22 days at 0.5 mg/kg to 50 mg/kg. Tumours were measured twice aweek and tumour volume calculated using the elliptical formula(pi/6×width×width×length). Data represent the geomean of tumour volumerelative to the tumour volume on day of randomisation. Error bars arethe 95% confidence interval (Graphpad Prism). This study demonstratedthat doses of 10 mg/kg and above gave tumour regression (FIG. 13).

Human ESR1 Mutant Breast Cancer Patient Derived Xenograft CTC174Anti-Tumour Efficacy in Mouse

To determine the effect of Example 17 on the growth of the ESR1 mutantpatient derived xenograft CTC174, female NSG mice were implanted withfragments of CTC174 in the mammary fat pad. CTC174 were derived fromcirculating tumour cells isolated from a patient with metastatic ER+breast cancer and have been shown to carry a D538G mutation in ESR1 at a0.33 allele frequency (Ladd, et al., Oncotarget, 2016, 7:54120-54136).Briefly, female ovariectomized NOD/SCID (Cg-Prkdcscid Il2rgtm1Wjl/SzJ)(NSG) mice (aged 6-7 weeks—The Jackson Laboratory) were implanted underrecoverable anaesthetic with a ˜50 mm³ fragment of a CTC174 xenograft inthe third mammary fat pad. When tumours reached ˜200 mm³ mice wererandomised into groups of 10 mice and started receiving drug treatment.Compounds were prepared in vehicle (40% Tetraethylene Glycol (v/v), 7.5%Captisol (w/v) in water for injection) and given orally in a volume of10 ml/kg once a day for 32 days at 0.8 mg/kg to 40 mg/kg. Tumours weremeasured twice a week and tumour volume calculated using the ellipticalformula (pi/6×width ×width×length). Data represent the geomean of tumourvolume relative to the tumour volume on day of randomisation. Error barsare the 95% confidence interval (Graphpad Prism). This studydemonstrated that doses of 10 mg/kg and above gave almost completetumour growth inhibition (FIG. 14).

Human ESR1 Mutant Breast Cancer Patient Derived Xenograft CTC174Anti-Tumour Drug Combination Efficacy in Mouse

To determine the effect of Example 17 on the growth of the ESR1 mutantpatient derived xenograft CTC174 in combination with the CDK4/6inhibitor palbociclib or the mTORC1/2 inhibitor vistusertib (AZD2014),female NSG mice were implanted with fragments of CTC174 in the mammaryfat pad. CTC174 were derived from circulating tumour cells isolated froma patient with metastatic ER+ breast cancer and have been shown to carrya D538G mutation in ESR1 at a 0.33 allele frequency (Ladd, et al.,Oncotarget, 2016, 7:54120-54136). Briefly, female NOD/SCID (Cg-PrkdcscidIl2rgtm1Wjl/SzJ) (NSG) mice (aged 6-7 weeks—The Jackson Laboratory) wereimplanted under recoverable anaesthetic with a ˜30 mm3 fragment of aCTC174 xenograft in the third mammary fat pad. When tumours reached ˜500mm3 mice were randomised into groups of 10 mice and started receivingdrug treatment. Ex17 was prepared in vehicle (40% Tetraethylene Glycol(v/v), 7.5% Captisol (w/v) in water for injection) and given orally in avolume of 10 ml/kg once a day for 23 days at 10 mg/kg. Palbociclib andvistusertib were prepared in vehicle (1% polysorbate 80). Palbociclibwas given orally in a volume of 10 ml/kg once a day for 23 days at 50mg/kg. Vistusertib was given orally in a volume of 10 ml/kg twice a dayin a 2 days on, 5 days off schedule for 23 days at 10 mg/kg. Vehicletreated group were dosed with 10 ml/kg 40% Tetraethylene Glycol (v/v),7.5% Captisol (w/v) in water for injection orally once a day for 23days. Tumours were measured twice a week and tumour volume calculatedusing the elliptical formula (pi/6×width×width×length). Data representthe geomean of tumour volume relative to the tumour volume on day ofrandomisation. Error bars are the 95% confidence interval (GraphpadPrism). This study demonstrated that Example 17 combined with eitherpalbociclib (FIG. 15) or with vistusertib (AZD2014) (FIG. 16) gave agreater effect than either agent dosed alone.

According to a further aspect of the specification there is provided apharmaceutical composition, which comprises a compound of the Formula(I), (IA), (IB), (IC) or (ID), or a pharmaceutically acceptable saltthereof, as defined hereinbefore in association with a pharmaceuticallyacceptable excipient.

Suitable pharmaceutically acceptable excipients for a tablet formulationinclude, for example, inert diluents, granulating and disintegratingagents, binding agents, lubricating agents, preservative agents andantioxidants. A further suitable pharmaceutically acceptable excipientmay be a chelating agent. Tablet formulations may be uncoated or coatedeither to modify their disintegration and the subsequent absorption ofthe active ingredient within the gastrointestinal tract, or to improvetheir stability and/or appearance, in either case, using conventionalcoating agents and procedures well known in the art.

Compositions for oral use may alternatively be in the form of hardgelatin capsules in which the active ingredient is mixed with an inertsolid diluent, or as soft gelatin capsules in which the activeingredient is mixed with water or an oil.

Aqueous suspensions generally contain the active ingredient in finelypowdered form together with one or more suspending agents, dispersing orwetting agents. The aqueous suspensions may also contain one or morepreservatives, anti-oxidants, colouring agents, flavouring agents,and/or sweetening agents.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil or in a mineral oil. The oily suspensions may alsocontain a thickening agent. Sweetening agents such as those set outabove, and flavouring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water generally contain the activeingredient together with a dispersing or wetting agent, suspending agentand one or more preservatives. Additional excipients such as sweetening,flavouring and colouring agents, may also be present.

The pharmaceutical compositions of the specification may also be in theform of oil-in-water emulsions. The oily phase may be a vegetable oil ora mineral oil or a mixture of any of these. The emulsions may alsocontain sweetening, flavouring and preservative agents.

Syrups and elixirs may be formulated with sweetening agents, and mayalso contain a demulcent, preservative, flavouring and/or colouringagent.

The pharmaceutical compositions may also be in the form of a sterileinjectable aqueous or oily suspension, which may be formulated accordingto known procedures using one or more of the appropriate dispersing orwetting agents and suspending agents, which have been mentioned above. Asterile injectable preparation may also be a sterile injectable solutionor suspension in a non-toxic parenterally-acceptable diluent or solventsystem.

Compositions for administration by inhalation may be in the form of aconventional pressurised aerosol arranged to dispense the activeingredient either as an aerosol containing finely divided solid orliquid droplets. Conventional aerosol propellants such as volatilefluorinated hydrocarbons or hydrocarbons may be used and the aerosoldevice is conveniently arranged to dispense a metered quantity of activeingredient. Dry powder inhalers may also be suitable.

For further information on formulation the reader is referred to Chapter25.2 in Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch;Chairman of Editorial Board), Pergamon Press 1990.

The amount of active ingredient that is combined with one or moreexcipients to produce a single dosage form will necessarily varydepending upon the host treated and the particular route ofadministration. For example, oral administration to humans willgenerally require, for example, from 1 mg to 2 g of active agent (moresuitably from 100 mg to 2 g, for example from 250 mg to 1.8 g, such asfrom 500 mg to 1.8 g, particularly from 500 mg to 1.5 g, convenientlyfrom 500 mg to 1 g) to be administered compounded with an appropriateand convenient amount of excipients which may vary from about 3 to about98 percent by weight of the total composition. It will be understoodthat, if a large dosage is required, multiple dosage forms may berequired, for example two or more tablets or capsules, with the dose ofactive ingredient divided conveniently between them. Typically, unitdosage forms will contain about 10 mg to 0.5 g of a compound of thisspecification, although a unit dosage form may contain up to 1 g.Conveniently, a single solid dosage form may contain between 1 and 300mg of active ingredient.

The size of the dose for therapeutic or prophylactic purposes ofcompounds of the present specification will naturally vary according tothe nature and severity of the disease state, the age and sex of theanimal or patient and the route of administration, according to wellknown principles of medicine.

In using compounds of the present specification for therapeutic orprophylactic purposes it will generally be administered so that a dailydose in the range, for example, 1 mg/kg to 100 mg/kg body weight isreceived, given if required in divided doses. In general, lower doseswill be administered when a parenteral route is employed. Thus, forexample, for intravenous administration, a dose in the range, forexample, 1 mg/kg to 25 mg/kg body weight will generally be used.Similarly, for administration by inhalation, a dose in the range, forexample, 1 mg/kg to 25 mg/kg body weight will be used. Oraladministration is however preferred, particularly in tablet form.

In one aspect of the specification, compounds of the presentspecification or pharmaceutically acceptable salts thereof, areadministered as tablets comprising 10 mg to 100 mg of the compound ofthe specification (or a pharmaceutically acceptable salt thereof),wherein one or more tablets are administered as required to achieve thedesired dose.

As stated above, it is known that signalling through ERα causestumourigenesis by one or more of the effects of mediating proliferationof cancer and other cells, mediating angiogenic events and mediating themotility, migration and invasiveness of cancer cells. We have found thatthe compounds of the present specification possess potent anti-tumouractivity which it is believed is obtained by way of antagonism anddown-regulation of ERα that is involved in the signal transduction stepswhich lead to the proliferation and survival of tumour cells and theinvasiveness and migratory ability of metastasising tumour cells.

Accordingly, the compounds of the present specification may be of valueas anti-tumour agents, in particular as selective inhibitors of theproliferation, survival, motility, dissemination and invasiveness ofmammalian cancer cells leading to inhibition of tumour growth andsurvival and to inhibition of metastatic tumour growth. Particularly,the compounds of the present specification may be of value asanti-proliferative and anti-invasive agents in the containment and/ortreatment of solid tumour disease. Particularly, the compounds of thepresent specification may be useful in the prevention or treatment ofthose tumours which are sensitive to inhibition of ERα and that areinvolved in the signal transduction steps which lead to theproliferation and survival of tumour cells and the migratory ability andinvasiveness of metastasising tumour cells. Further, the compounds ofthe present specification may be useful in the prevention or treatmentof those tumours which are mediated alone or in part by antagonism anddown-regulation of ERα, i.e. the compounds may be used to produce an ERαinhibitory effect in a warm-blooded animal in need of such treatment.

According to a further aspect of the specification there is provided acompound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore foruse as a medicament in a warm-blooded animal such as man.

According to a further aspect of the specification, there is provided acompound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore foruse in the production of an anti-proliferative effect in a warm-bloodedanimal such as man.

According to a further aspect of the specification there is provided acompound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore foruse in a warm-blooded animal such as man as an anti-invasive agent inthe containment and/or treatment of solid tumour disease.

According to a further aspect of the specification, there is providedthe use of a compound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore, forthe production of an anti-proliferative effect in a warm-blooded animalsuch as man.

According to a further aspect of the specification there is provided theuse of a compound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore, inthe manufacture of a medicament for use in the production of ananti-proliferative effect in a warm-blooded animal such as man.

According to a further aspect of the specification there is provided theuse of a compound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore, inthe manufacture of a medicament for use in a warm-blooded animal such asman as an anti-invasive agent in the containment and/or treatment ofsolid tumour disease.

According to a further aspect of the specification there is provided amethod for producing an anti-proliferative effect in a warm-bloodedanimal, such as man, in need of such treatment which comprisesadministering to said animal an effective amount of a compound of theFormula (I), (IA), (IB), (IC) or (ID), or a pharmaceutically acceptablesalt thereof, as defined hereinbefore.

According to a further aspect of the specification there is provided amethod for producing an anti-invasive effect by the containment and/ortreatment of solid tumour disease in a warm-blooded animal, such as man,in need of such treatment which comprises administering to said animalan effective amount of a compound of the Formula (I), (IA), (IB), (IC)or (ID), or a pharmaceutically acceptable salt thereof, as definedhereinbefore.

According to a further aspect of the specification, there is provided acompound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore, foruse in the prevention or treatment of cancer in a warm-blooded animalsuch as man.

According to a further aspect of the specification there is provided theuse of a compound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore in themanufacture of a medicament for use in the prevention or treatment ofcancer in a warm-blooded animal such as man.

According to a further aspect of the specification there is provided amethod for the prevention or treatment of cancer in a warm-bloodedanimal, such as man, in need of such treatment which comprisesadministering to said animal an effective amount of a compound of theFormula (I), (IA), (IB), (IC) or (ID), or a pharmaceutically acceptablesalt thereof, as defined hereinbefore.

According to a further aspect of the specification, there is provided acompound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore foruse in the prevention or treatment of solid tumour disease in awarm-blooded animal such as man.

According to a further aspect of the specification there is provided theuse of a compound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore, inthe manufacture of a medicament for use in the prevention or treatmentof solid tumour disease in a warm-blooded animal such as man.

According to a further aspect of the specification there is provided amethod for the prevention or treatment of solid tumour disease in awarm-blooded animal, such as man, in need of such treatment whichcomprises administering to said animal an effective amount of a compoundof the Formula (I), (IA), (IB), (IC) or (ID), or a pharmaceuticallyacceptable salt thereof, as defined hereinbefore.

According to a further aspect of the specification there is provided acompound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore, foruse in the prevention or treatment of those tumours which are sensitiveto inhibition of ERα that are involved in the signal transduction stepswhich lead to the proliferation, survival, invasiveness and migratoryability of tumour cells.

According to a further aspect of the specification there is provided theuse of a compound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore, inthe manufacture of a medicament for use in the prevention or treatmentof those tumours which are sensitive to inhibition of ERα that areinvolved in the signal transduction steps which lead to theproliferation, survival, invasiveness and migratory ability of tumourcells.

According to a further aspect of the specification there is provided amethod for the prevention or treatment of those tumours which aresensitive to inhibition of ERα that are involved in the signaltransduction steps which lead to the proliferation, survival,invasiveness and migratory ability of tumour cells which comprisesadministering to said animal an effective amount of a compound of theFormula (I), (IA), (IB), (IC) or (ID), or a pharmaceutically acceptablesalt thereof, as defined hereinbefore.

According to a further aspect of the specification there is provided acompound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore foruse in providing an inhibitory effect on ERα.

According to a further aspect of the specification there is provided theuse of a compound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore in themanufacture of a medicament for use in providing an inhibitory effect onERα.

According to a further aspect of the specification there is alsoprovided a method for providing an inhibitory effect on ERα whichcomprises administering an effective amount of a compound of the Formula(I), (IA), (IB), (IC) or (ID), or a pharmaceutically acceptable saltthereof, as defined hereinbefore.

According to a further aspect of the specification there is provided acompound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore, foruse in providing a selective inhibitory effect on ERα.

According to a further aspect of the specification there is provided theuse of a compound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore, inthe manufacture of a medicament for use in providing a selectiveinhibitory effect on ERα.

According to a further aspect of the specification there is alsoprovided a method for providing a selective inhibitory effect on ERαwhich comprises administering an effective amount of a compound of theFormula (I), (IA), (IB), (IC) or (ID), or a pharmaceutically acceptablesalt thereof, as defined hereinbefore.

Described herein are compounds that can bind to ERα ligand bindingdomain and are selective estrogen receptor degraders. In biochemical andcell based assays the compounds of the present specification are shownto be potent estrogen receptor binders and reduce cellular levels of ERαand may therefore be useful in the treatment of estrogen sensitivediseases or conditions (including diseases that have developedresistance to endocrine therapies), i.e. for use in the treatment ofcancer of the breast and gynaecological cancers (including endometrial,ovarian and cervical) and cancers expressing ERα mutated proteins whichmay be de novo mutations or have arisen as a result of treatment with aprior endocrine therapy such as an aromatase inhibitor.

According to a further aspect of the specification there is provided acompound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore, foruse in the treatment of breast or gynaecological cancers.

According to a further aspect of the specification there is provided acompound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore, foruse in the treatment of cancer of the breast, endometrium, ovary orcervix.

According to a further aspect of the specification there is provided acompound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore, foruse in the treatment of cancer of the breast.

According to a further aspect of the specification there is provided acompound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore, foruse in the treatment of cancer of the breast, wherein the cancer hasdeveloped resistance to one or more other endocrine therapies.

According to a further aspect of the specification there is provided amethod for treating breast or gynaecological cancers, which comprisesadministering an effective amount of a compound of the Formula (I),(IA), (IB), (IC) or (ID), or a pharmaceutically acceptable salt thereof,as defined hereinbefore.

According to a further aspect of the specification there is provided amethod for treating cancer of the breast, endometrium, ovary or cervix,which comprises administering an effective amount of a compound of theFormula (I), (IA), (IB), (IC) or (ID), or a pharmaceutically acceptablesalt thereof, as defined hereinbefore.

According to a further aspect of the specification there is provided amethod for treating breast cancer, which comprises administering aneffective amount of a compound of the Formula (I), (IA), (IB), (IC) or(ID), or a pharmaceutically acceptable salt thereof, as definedhereinbefore.

According to a further aspect of the specification there is provided amethod for treating breast cancer, wherein the cancer has developedresistance to one or more other endocrine therapies, which comprisesadministering an effective amount of a compound of the Formula (I),(IA), (IB), (IC) or (ID), or a pharmaceutically acceptable salt thereof,as defined hereinbefore.

According to a further aspect of the specification there is provided theuse of a compound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore, inthe manufacture of a medicament for use in the treatment of breast orgynaecological cancers.

According to a further aspect of the specification there is provided theuse of a compound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore, inthe manufacture of a medicament for use in the treatment of cancer ofthe breast, endometrium, ovary or cervix.

According to a further aspect of the specification there is provided theuse of a compound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore, inthe manufacture of a medicament for use in the treatment of breastcancer.

According to a further aspect of the specification there is provided theuse of a compound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore, inthe manufacture of a medicament for use in the treatment of breastcancer, wherein the cancer has developed resistance to one or more otherendocrine therapies.

In one feature of the specification, the cancer to be treated is breastcancer. In a further aspect of this feature, the breast cancer isEstrogen Receptor+ve (ER+ve). In one embodiment of this aspect, thecompound of Formula (I), (IA), (IB), (IC) or (ID), is dosed incombination with another anticancer agent, such as an anti-hormonalagent as defined herein.

According to a further aspect of the specification there is provided acompound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined hereinbefore, foruse in the treatment of ER+ve breast cancer.

According to a further aspect of the specification there is provided amethod for treating ER+ve breast cancer, which comprises administeringan effective amount of a compound of the Formula (I), (IA), (IB), (IC)or (ID), or a pharmaceutically acceptable salt thereof, as definedhereinbefore.

According to a further aspect of the specification there is provided theuse of a compound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, as defined herein before inthe manufacture of a medicament for use in the treatment of ER+ve breastcancer.

As stated hereinbefore, the in-vivo effects of a compound of the Formula(I), (IA), (IB), (IC) or (ID) may be exerted in part by one or moremetabolites that are formed within the human or animal body afteradministration of a compound of the Formula (I), (IA), (IB), (IC) or(ID).

The present specification therefore also contemplates a method forinhibiting ER-α in a patient, comprising administering to a patient anamount of a compound of Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, effective in inhibiting ER-αin the patient.

The present specification therefore also contemplates a method forinhibiting ER-α in a patient, comprising administering to a patient anamount of a compound of Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, effective in inhibiting ER-αin the patient.

The anti-cancer treatment defined herein may be applied as a soletherapy or may involve, in addition to the compounds of thespecification, conventional surgery or radiotherapy or chemotherapy.Such chemotherapy may include one or more of the following categories ofanti-tumour agents:—

(i) other antiproliferative/antineoplastic drugs and combinationsthereof, as used in medical oncology, such as alkylating agents (forexample cis-platin, oxaliplatin, carboplatin, cyclophosphamide, nitrogenmustard, melphalan, chlorambucil, busulphan, temozolamide andnitrosoureas); antimetabolites (for example gemcitabine and antifolatessuch as fluoropyrimidines like 5-fluorouracil and tegafur, raltitrexed,methotrexate, cytosine arabinoside, and hydroxyurea); antitumourantibiotics (for example anthracyclines like adriamycin, bleomycin,doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C,dactinomycin and mithramycin); antimitotic agents (for example vincaalkaloids like vincristine, vinblastine, vindesine and vinorelbine andtaxoids like taxol and taxotere and polokinase inhibitors); andtopoisomerase inhibitors (for example epipodophyllotoxins like etoposideand teniposide, amsacrine, topotecan and camptothecin);(ii) antihormonal agents such as antioestrogens (for example tamoxifen,fulvestrant, toremifene, raloxifene, droloxifene and iodoxyfene),progestogens (for example megestrol acetate), aromatase inhibitors (forexample as anastrozole, letrozole, vorazole and exemestane);(iii) inhibitors of growth factor function and their downstreamsignalling pathways: included are Ab modulators of any growth factor orgrowth factor receptor targets, reviewed by Stern et al. CriticalReviews in Oncology/Haematology, 2005, 54, pp 11-29); also included aresmall molecule inhibitors of such targets, for example kinaseinhibitors—examples include the anti-erbB2 antibody trastuzumab[Herceptin™], the anti-EGFR antibody panitumumab, the anti-EGFR antibodycetuximab [Erbitux, C225] and tyrosine kinase inhibitors includinginhibitors of the erbB receptor family, such as epidermal growth factorfamily receptor (EGFR/erbB1) tyrosine kinase inhibitors such asgefitinib or erlotinib, erbB2 tyrosine kinase inhibitors such aslapatinib, and mixed erb1/2 inhibitors such as afatanib; similarstrategies are available for other classes of growth factors and theirreceptors, for example inhibitors of the hepatocyte growth factor familyor their receptors including c-met and ron; inhibitors of the insulinand insulin growth factor family or their receptors (IGFR, IR)inhibitors of the platelet-derived growth factor family or theirreceptors (PDGFR), and inhibitors of signalling mediated by otherreceptor tyrosine kinases such as c-kit, AnLK, and CSF-1R; also includedare modulators which target signalling proteins in the PI3-kinasesignalling pathway, for example, inhibitors of PI3-kinase isoforms suchas PI3K-α/β/γ and ser/thr kinases such as AKT, mTOR (such as AZD2014),PDK, SGK, PI4K or PIP5K; also included are inhibitors ofserine/threonine kinases not listed above, for example raf inhibitorssuch as vemurafenib, MEK inhibitors such as selumetinib (AZD6244), Ablinhibitors such as imatinib or nilotinib, Btk inhibitors such asibrutinib, Syk inhibitors such as fostamatinib, aurora kinase inhibitors(for example AZD1152), inhibitors of other ser/thr kinases such as JAKs,STATs and IRAK4, and cyclin dependent kinase inhibitors for exampleinhibitors of CDK1, CDK7, CDK9 and CDK4/6 such as palbociclib;iv) modulators of DNA damage signalling pathways, for example PARPinhibitors (e.g. Olaparib), ATR inhibitors or ATM inhibitors;v) modulators of apoptotic and cell death pathways such as Bcl familymodulators (e.g. ABT-263/Navitoclax, ABT-199);(vi) antiangiogenic agents such as those which inhibit the effects ofvascular endothelial growth factor, [for example the anti-vascularendothelial cell growth factor antibody bevacizumab (Avastin™) and forexample, a VEGF receptor tyrosine kinase inhibitor such as sorafenib,axitinib, pazopanib, sunitinib and vandetanib (and compounds that workby other mechanisms (for example linomide, inhibitors of integrin αvβ3function and angiostatin)];(vii) vascular damaging agents, such as Combretastatin A4;(viii) anti-invasion agents, for example c-Src kinase family inhibitorslike (dasatinib, J. Med. Chem., 2004, 47, 6658-6661) and bosutinib(SKI-606), and metalloproteinase inhibitors like marimastat, inhibitorsof urokinase plasminogen activator receptor function or antibodies toHeparanase];(ix) immunotherapy approaches, including for example ex-vivo and in-vivoapproaches to increase the immunogenicity of patient tumour cells, suchas transfection with cytokines such as interleukin 2, interleukin 4 orgranulocyte-macrophage colony stimulating factor, approaches to decreaseT-cell anergy, approaches using transfected immune cells such ascytokine-transfected dendritic cells, approaches usingcytokine-transfected tumour cell lines and approaches usinganti-idiotypic antibodies. Specific examples include monoclonalantibodies targeting PD-1 (e.g. BMS-936558) or CTLA4 (e.g. ipilimumaband tremelimumab);(x) Antisense or RNAi based therapies, for example those which aredirected to the targets listed.(xi) gene therapy approaches, including for example approaches toreplace aberrant genes such as aberrant p53 or aberrant BRCA1 or BRCA2,GDEPT (gene-directed enzyme pro-drug therapy) approaches such as thoseusing cytosine deaminase, thymidine kinase or a bacterial nitroreductaseenzyme and approaches to increase patient tolerance to chemotherapy orradiotherapy such as multi-drug resistance gene therapy.

Accordingly, in one embodiment there is provided a compound of Formula(I), (IA), (IB), (IC) or (ID), or a pharmaceutically acceptable saltthereof, and an additional anti-tumour substance for the conjointtreatment of cancer.

According to this aspect of the specification there is provided acombination suitable for use in the treatment of cancer comprising acompound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof and another anti-tumour agent,in particular any one of the anti tumour agents listed under (i)-(xi)above. In particular, the anti-tumour agent listed under (i)-(xi) aboveis the standard of care for the specific cancer to be treated; theperson skilled in the art will understand the meaning of “standard ofcare”.

Therefore in a further aspect of the specification there is provided acompound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, in combination with anotheranti-tumour agent, in particular an anti-tumour agent selected from onelisted under (i)-(xi) herein above.

In a further aspect of the specification there is provided a compound ofthe Formula (I), (IA), (IB), (IC) or (ID), or a pharmaceuticallyacceptable salt thereof, in combination with another anti-tumour agent,in particular an anti-tumour agent selected from one listed under (i)above.

In a further aspect of the specification there is provided a compound ofthe Formula (I), (IA), (IB), (IC) or (ID), or a pharmaceuticallyacceptable salt thereof, and any one of the anti-tumour agents listedunder (i) above.

In a further aspect of the specification there is provided a combinationsuitable for use in the treatment of cancer comprising a compound of theFormula (I), (IA), (IB), (IC) or (ID), or a pharmaceutically acceptablesalt thereof, and a taxoid, such as for example taxol or taxotere,conveniently taxotere.

In a further aspect of the specification there is provided a compound ofthe Formula (I), (IA), (IB), (IC) or (ID), or a pharmaceuticallyacceptable salt thereof, in combination with another anti-tumour agent,in particular an anti-tumour agent selected from one listed under (ii)herein above.

In a further aspect of the specification there is provided a combinationsuitable for use in the treatment of cancer comprising a compound of theFormula (I), (IA), (IB), (IC) or (ID), or a pharmaceutically acceptablesalt thereof, and any one of the antihormonal agents listed under (ii)above, for example any one of the anti-oestrogens listed in (ii) above,or for example an aromatase inhibitor listed in (ii) above.

In a further aspect of the specification there is provided a combinationsuitable for use in the treatment of cancer comprising a compound of theFormula (I), (IA), (IB), (IC) or (ID), or a pharmaceutically acceptablesalt thereof, and an mTOR inhibitor, such as AZD2014.

In a further aspect of the specification there is provided a combinationsuitable for use in the treatment of cancer comprising a compound of theFormula (I), (IA), (IB), (IC) or (ID), or a pharmaceutically acceptablesalt thereof, and a PI3Kα-inhibitor, such as the compound1-(4-(5-(5-amino-6-(5-tert-butyl-1,3,4-oxadiazol-2-yl)pyrazin-2-yl)-1-ethyl-1H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-hydroxypropan-1-one,or a pharmaceutically-acceptable salt thereof.

In a further aspect of the specification there is provided a combinationsuitable for use in the treatment of cancer comprising a compound of theFormula (I), (IA), (IB), (IC) or (ID), or a pharmaceutically acceptablesalt thereof, and a CDK4/6 inhibitor, such as palbociclib.

In one aspect the above combination of a compound of the Formula (I),(IA), (IB), (IC) or (ID), or a pharmaceutically acceptable salt thereof,with an anti-tumour agent listed in (ii) above, or an mTOR inhibitor(such as AZD2014), or a PI3K-α inhibitor (such as the compound1-(4-(5-(5-amino-6-(5-tert-butyl-1,3,4-oxadiazol-2-yl)pyrazin-2-yl)-1-ethyl-1H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-hydroxypropan-1-one)or a CDK4/6 inhibitor (such as palbociclib), is suitable for use in thetreatment of breast or gynaecological cancers, such as cancer of thebreast, endometrium, ovary or cervix, particularly breast cancer, suchas ER+ve breast cancer.

Herein, where the term “combination” is used it is to be understood thatthis refers to simultaneous, separate or sequential administration. Inone aspect of the specification “combination” refers to simultaneousadministration. In another aspect of the specification “combination”refers to separate administration. In a further aspect of thespecification “combination” refers to sequential administration. Wherethe administration is sequential or separate, the delay in administeringthe second component should not be such as to lose the beneficial effectof the combination. Where a combination of two or more components isadministered separately or sequential, it will be understood that thedosage regime for each component may be different to and independent ofthe other components. Conveniently, the compounds of the presentspecification are dosed once daily.

According to a further aspect of the specification there is provided apharmaceutical composition which comprises a compound of Formula (I),(IA), (IB), (IC) or (ID), or a pharmaceutically acceptable salt thereofin combination with an anti-tumour agent selected from one listed under(i)-(xi) herein above, in association with a pharmaceutically acceptableexcipient.

According to a further aspect of the specification there is provided apharmaceutical composition which comprises a compound of Formula (I),(IA), (IB), (IC) or (ID), or a pharmaceutically acceptable salt thereofin combination with any one of antihormonal agents listed under (ii)above, for example any one of the anti-oestrogens listed in (ii) above,or for example an aromatase inhibitor listed in (ii) above inassociation with a pharmaceutically acceptable excipient.

In a further aspect of the specification there is provided apharmaceutical composition comprising a compound of the Formula (I),(IA), (IB), (IC) or (ID), or a pharmaceutically acceptable salt thereof,and an mTOR inhibitor, such as AZD2014, in association with apharmaceutically acceptable excipient.

In a further aspect of the specification there is provided apharmaceutical composition comprising a compound of the Formula (I),(IA), (IB), (IC) or (ID), or a pharmaceutically acceptable salt thereof,and a PI3Kα-inhibitor, such as the compound1-(4-(5-(5-amino-6-(5-tert-butyl-1,3,4-oxadiazol-2-yl)pyrazin-2-yl)-1-ethyl-1H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-hydroxypropan-1-one,in association with a pharmaceutically acceptable excipient.

In a further aspect of the specification there is provided apharmaceutical composition comprising a compound of the Formula (I),(IA), (IB), (IC) or (ID), or a pharmaceutically acceptable salt thereof,and a CDK4/6 inhibitor (such as palbociclib) in association with apharmaceutically acceptable excipient.

According to a further aspect of the specification there is provided apharmaceutical composition which comprises a compound of the Formula(I), (IA), (IB), (IC) or (ID), or a pharmaceutically acceptable saltthereof, in combination with an anti-tumour agent selected from onelisted under (i)-(xi) herein above, in association with apharmaceutically acceptable excipient for use in treating cancer.

According to a further aspect of the specification there is provided apharmaceutical composition which comprises a compound of the Formula(I), (IA), (IB), (IC) or (ID), or a pharmaceutically acceptable saltthereof, in combination with any one of antihormonal agents listed under(ii) above, for example any one of the anti-oestrogens listed in (ii)above, or for example an aromatase inhibitor listed in (ii) above inassociation with a pharmaceutically acceptable excipient for use intreating cancer.

In a further aspect of the specification there is provided apharmaceutical composition comprising a compound of the Formula (I),(IA), (IB), (IC) or (ID), or a pharmaceutically acceptable salt thereof,and an mTOR inhibitor, such as AZD2014, in association with apharmaceutically acceptable excipient for use in treating cancer.

In a further aspect of the specification there is provided apharmaceutical composition comprising a compound of the Formula (I),(IA), (IB), (IC) or (ID), or a pharmaceutically acceptable salt thereof,and a PI3Kα-inhibitor, such as the compound1-(4-(5-(5-amino-6-(5-tert-butyl-1,3,4-oxadiazol-2-yl)pyrazin-2-yl)-1-ethyl-1H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-hydroxypropan-1-one,in association with a pharmaceutically acceptable excipient for use intreating cancer.

In a further aspect of the specification there is provided apharmaceutical composition comprising a compound of the Formula (I),(IA), (IB), (IC) or (ID), or a pharmaceutically acceptable salt thereof,and a CDK4/6 inhibitor (such as palbociclib) in association with apharmaceutically acceptable excipient for use in treating cancer.

In one aspect the above pharmaceutical compositions of a compound of theFormula (I), (IA), (IB), (IC) or (ID), or a pharmaceutically acceptablesalt thereof, with an anti-tumour agent listed in (ii) above, or an mTORinhibitor (such as AZD2014), or a PI3K-α inhibitor (such as the compound1-(4-(5-(5-amino-6-(5-tert-butyl-1,3,4-oxadiazol-2-yl)pyrazin-2-yl)-1-ethyl-1H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-hydroxypropan-1-one)or a CDK4/6 inhibitor (such as palbociclib), is suitable for use in thetreatment of breast or gynaecological cancers, such as cancer of thebreast, endometrium, ovary or cervix, particularly breast cancer, suchas ER+ve breast cancer.

According to another feature of the specification there is provided theuse of a compound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof in combination with ananti-tumour agent selected from one listed under (i)-(xi) herein above,in the manufacture of a medicament for use in the treatment of cancer ina warm-blooded animal, such as man.

According to a further aspect of the specification there is provided theuse of a compound of Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof in combination with any one ofantihormonal agents listed under (ii) above, for example any one of theanti-oestrogens listed in (ii) above, or for example an aromataseinhibitor listed in (ii) above in the manufacture of a medicament foruse in the treatment of cancer in a warm-blooded animal, such as man.

In a further aspect of the specification there is provided the use of acompound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, in combination with an mTORinhibitor, such as AZD2014, in the manufacture of a medicament for usein the treatment of cancer in a warm-blooded animal, such as man.

In a further aspect of the specification there is provided the use of acompound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, in combination with aPI3Kα-inhibitor, such as the compound1-(4-(5-(5-amino-6-(5-tert-butyl-1,3,4-oxadiazol-2-yl)pyrazin-2-yl)-1-ethyl-1H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-hydroxypropan-1-one,in the manufacture of a medicament for use in the treatment of cancer ina warm-blooded animal, such as man.

In a further aspect of the specification there is provided the use acompound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, in combination with a CDK4/6inhibitor (such as palbociclib) in the manufacture of a medicament foruse in the treatment of cancer in a warm-blooded animal, such as man.

In one aspect the above uses of a compound of the Formula (I), (IA),(IB), (IC) or (ID), or a pharmaceutically acceptable salt thereof, incombination with an anti-tumour agent listed in (ii) above, or an mTORinhibitor (such as AZD2014), or a PI3K-α inhibitor (such as the compound1-(4-(5-(5-amino-6-(5-tert-butyl-1,3,4-oxadiazol-2-yl)pyrazin-2-yl)-1-ethyl-1H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-hydroxypropan-1-one)or a CDK4/6 inhibitor (such as palbociclib), is suitable for use in themanufacture of a medicament for the treatment of breast orgynaecological cancers, such as cancer of the breast, endometrium, ovaryor cervix, particularly breast cancer, such as ER+ve breast cancer.

Therefore in an additional feature of the specification, there isprovided a method of treating cancer in a warm-blooded animal, such asman, in need of such treatment which comprises administering to saidanimal an effective amount of a compound of the Formula (I), or apharmaceutically acceptable salt thereof, in combination with ananti-tumour agent selected from one listed under (i)-(xi) herein above.

According to a further aspect of the specification there is provided amethod of treating cancer in a warm-blooded animal, such as man, in needof such treatment which comprises administering to said animal aneffective amount of a compound of Formula (I), (IA), (IB), (IC) or (ID),or a pharmaceutically acceptable salt thereof in combination with anyone of antihormonal agents listed under (ii) above, for example any oneof the anti-oestrogens listed in (ii) above, or for example an aromataseinhibitor listed in (ii) above.

In a further aspect of the specification there is provided a method oftreating cancer in a warm-blooded animal, such as man, in need of suchtreatment which comprises administering to said animal an effectiveamount of a compound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, in combination with an mTORinhibitor, such as AZD2014.

In a further aspect of the specification there provided a method oftreating cancer in a warm-blooded animal, such as man, in need of suchtreatment which comprises administering to said animal an effectiveamount of a compound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, in combination with aPI3Kα-inhibitor, such as the compound1-(4-(5-(5-amino-6-(5-tert-butyl-1,3,4-oxadiazol-2-yl)pyrazin-2-yl)-1-ethyl-1H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-hydroxypropan-1-one.

In a further aspect of the specification there is provided a method oftreating cancer in a warm-blooded animal, such as man, in need of suchtreatment which comprises administering to said animal an effectiveamount of a compound of the Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof, in combination with a CDK4/6inhibitor (such as palbociclib).

In one aspect the above combinations, pharmaceutical compositions, usesand methods of treating cancer, are methods for the treatment of breastor gynaecological cancers, such as cancer of the breast, endometrium,ovary or cervix, particularly breast cancer, such as ER+ve breastcancer.

According to a further aspect of the present specification there isprovided a kit comprising a compound of Formula (I), (IA), (IB), (IC) or(ID), or a pharmaceutically acceptable salt thereof in combination withan anti-tumour agent selected from one listed under (i)-(xi) hereinabove.

According to a further aspect of the present specification there isprovided a kit comprising a compound of Formula (I), (IA), (IB), (IC) or(ID), or a pharmaceutically acceptable salt thereof in combination withan anti-tumour agent selected from one listed under (i) or (ii) hereinabove.

According to a further aspect of the present specification there isprovided a kit comprising:

-   -   a) a compound of Formula (I), (IA), (IB), (IC) or (ID), or a        pharmaceutically acceptable salt thereof in a first unit dosage        form;    -   b) an anti-tumour agent selected from one listed under (i)-(xi)        herein above in a second unit dosage form; and    -   c) container means for containing said first and second dosage        forms.

According to a further aspect of the present specification there isprovided a kit comprising:

-   -   a) a compound of Formula (I), (IA), (IB), (IC) or (ID), or a        pharmaceutically acceptable salt thereof in a first unit dosage        form;    -   b) an anti-tumour agent selected from one listed under (i)-(ii)        herein above in a second unit dosage form; and    -   c) container means for containing said first and second dosage        forms.

According to a further aspect of the present specification there isprovided a kit comprising:

-   -   a) a compound of the Formula (I), (IA), (IB), (IC) or (ID), or a        pharmaceutically acceptable salt thereof, in a first unit dosage        form;    -   b) an anti-tumour agent selected from an anti-tumour agent        listed in (ii) above, an mTOR inhibitor (such as AZD2014), a        PI3Kα-inhibitor, such as the compound        1-(4-(5-(5-amino-6-(5-tert-butyl-1,3,4-oxadiazol-2-yl)pyrazin-2-yl)-1-ethyl-1H-1,2,4-triazol-3-yl)piperidin-1-yl)-3-hydroxypropan-1-one,        and a CDK4/6 inhibitor, such as palbociclib, in a second unit        dosage form; and c) container means for containing said first        and second dosage forms.

Combination therapy as described above may be added on top of standardof care therapy typically carried out according to its usual prescribingschedule.

Although the compounds of the Formula (I), (IA), (IB), (IC) or (ID), areprimarily of value as therapeutic agents for use in warm-blooded animals(including man), they are also useful whenever it is required to inhibitER-α. Thus, they are useful as pharmacological standards for use in thedevelopment of new biological tests and in the search for newpharmacological agents.

Personalised Healthcare

Another aspect of the present specification is based on identifying alink between the status of the gene encoding ERα and potentialsusceptibility to treatment with a compound of Formula (I), (IA), (IB),(IC) or (ID). In particular, ERα gene status may indicate that a patientis less likely to respond to exisiting hormone therapy (such asaromatase inhibitors), in part at least because some ERα mutations arethough to arise as resistance mechanisms to existing treatments. A SERD,particularly a SERD which can be administered orally in potentiallylarger doses without excessive inconvenince, may then advantageously beused to treat patentients with ERα mutations who may be resistant toother therapies. This therefore provides opportunities, methods andtools for selecting patients for treatment with a compound of Formula(I), (IA), (IB), (IC) or (ID), particularly cancer patients. The presentspecification relates to patient selection tools and methods (includingpersonalised medicine). The selection is based on whether the tumourcells to be treated possess wild-type or mutant ERα gene. The ERα genestatus could therefore be used as a biomarker to indicate that selectingtreatment with a SERD may be advantageous. For the avoidance of doubt,compounds of the Formula (I), (IA), (IB), (IC) or (ID), as describedherein, are thought to be similarly active against wild-type and mutantERα genes, at least those mutations in ERα gene identified at the dateof filing this application.

There is a clear need for biomarkers that will enrich for or selectpatients whose tumours will respond to treatment with a SERD, such as acompound of Formula (I), (IA), (IB), (IC) or (ID). Patient selectionbiomarkers that identify the patients most likely to respond to oneagent over another are ideal in the treatment of cancer, since theyreduce the unnecessary treatment of patients with non-responding tumoursto the potential side effects of such agents.

A biomarker can be described as “a characteristic that is objectivelymeasured and evaluated as an indicator of normal biologic processes,pathogenic processes, or pharmacologic responses to a therapeuticintervention”. A biomarker is any identifiable and measurable indicatorassociated with a particular condition or disease where there is acorrelation between the presence or level of the biomarker and someaspect of the condition or disease (including the presence of, the levelor changing level of, the type of, the stage of, the susceptibility tothe condition or disease, or the responsiveness to a drug used fortreating the condition or disease). The correlation may be qualitative,quantitative, or both qualitative and quantitative. Typically abiomarker is a compound, compound fragment or group of compounds. Suchcompounds may be any compounds found in or produced by an organism,including proteins (and peptides), nucleic acids and other compounds.

Biomarkers may have a predictive power, and as such may be used topredict or detect the presence, level, type or stage of particularconditions or diseases (including the presence or level of particularmicroorganisms or toxins), the susceptibility (including geneticsusceptibility) to particular conditions or diseases, or the response toparticular treatments (including drug treatments). It is thought thatbiomarkers will play an increasingly important role in the future ofdrug discovery and development, by improving the efficiency of researchand development programs. Biomarkers can be used as diagnostic agents,monitors of disease progression, monitors of treatment and predictors ofclinical outcome. For example, various biomarker research projects areattempting to identify markers of specific cancers and of specificcardiovascular and immunological diseases. It is believed that thedevelopment of new validated biomarkers will lead both to significantreductions in healthcare and drug development costs and to significantimprovements in treatment for a wide variety of diseases and conditions.

In order to optimally design clinical trials and to gain the mostinformation from these trials, a biomarker may be required. The markermay be measurable in surrogate and tumour tissues. Ideally these markerswill also correlate with efficacy and thus could ultimately be used forpatient selection.

Thus, the technical problem underlying this aspect of the presentspecification is the identification of means for stratification ofpatients for treatment with a compound of Formula (I). The technicalproblem is solved by provision of the embodiments characterized in theclaims and/or description herein.

Tumours which contain wild type ERα are believed to be susceptible totreatment with a compound of Formula (I), (IA), (IB), (IC) or (ID), forexample as a first-line treatment. Tumours may also respond to treatmentwith a compound of Formula (I), (IA),

(IB), (IC) or (ID) as a second-line, third-line or subsequent therapyand this may be useful, in particular, where the tumours contain mutantERα and may thus be resistant to existing therapies such as AIs. Ahigher dosage of a compound of Formula (I), (IA), (IB), (IC) or (ID) maybe required in the resistant setting than in wild type tumours).

The specification provides a method of determining sensitivity of cellsto a compound of Formula (I), (IA), (IB), (IC) or (ID). The methodcomprises determining the status of ERα gene in said cells. A cell isdefined as sensitive to a compound of Formula (I), (IA), (IB), (IC) or(ID) if it inhibits the increase in cell number in a cell growth assay(either through inhibition of cell proliferation and/or throughincreased cell death). Methods of the specification are useful forpredicting which cells are more likely to respond to a compound ofFormula (I), (IA), (IB), (IC) or (ID) by growth inhibition.

A sample “representative of the tumour” can be the actual tumour sampleisolated, or may be a sample that has been further processed, e.g. asample of PCR amplified nucleic acid from the tumour sample.

Definitions

In this Personalised Healthcare section:

“Allele” refers to a particular form of a genetic locus, distinguishedfrom other forms by its particular nucleotide or amino acid sequence.

“Amplification reactions” are nucleic acid reactions which result inspecific amplification of target nucleic acids over non-target nucleicacids. The polymerase chain reaction (PCR) is a well known amplificationreaction.

“Cancer” is used herein to refer to neoplastic growth arising fromcellular transformation to a neoplastic phenotype. Such cellulartransformation often involves genetic mutation.

“Gene” is a segment of DNA that contains all the information for theregulated biosynthesis of an RNA product, including a promoter, exons,introns, and other sequence elements which may be located within 5′ or3′ flanking regions (not within the transcribed portions of the gene)that control expression.

“Gene status” refers to whether the gene is wild type or not (i.e.mutant).

“Label” refers to a composition capable of producing a detectable signalindicative of the presence of the target polynucleotide in an assaysample. Suitable labels include radioisotopes, nucleotide chromophores,enzymes, substrates, fluorescent molecules, chemiluminescent moieties,magnetic particles, bioluminescent moieties, and the like. As such, alabel is any composition detectable by spectroscopic, photochemical,biochemical, immunochemical, electrical, optical or chemical means.

“Non-synonymous variation” refers to a variation (variance) in oroverlapping the coding sequence of a gene that result in the productionof a distinct (altered) polypeptide sequence. These variations may ormay not affect protein function and include missense variants (resultingin substitution of one amino acid for another), nonsense variants(resulting in a truncated polypeptide due to generation of a prematurestop codon) and insertion/deletion variants.

“Synonymous variation” refers to a variation (variance) in the codingsequence of a gene that does not affect sequence of the encodedpolypeptide. These variations may affect protein function indirectly(for example by altering expression of the gene), but, in the absence ofevidence to the contrary, are generally assumed to be innocuous.

“Nucleic acid” refers to single stranded or double stranded DNA and RNAmolecules including natural nucleic acids found in nature and/ormodified, artificial nucleic acids having modified backbones or bases,as are known in the art.

“Primer” refers to a single stranded DNA oligonucleotide sequencecapable of acting as a point of initiation for synthesis of a primerextension product which is complementary to the nucleic acid strand tobe copied. The length and sequence of the primer must be such that theyare able to prime the synthesis of extension products. A typical primercontains at least about 7 nucleotides in length of a sequencesubstantially complementary to the target sequence, but somewhat longerprimers are preferred. Usually primers contain about 15-26 nucleotides,but longer or shorter primers may also be employed.

“Polymorphic site” is a position within a locus at which at least twoalternative sequences are found in a population.

“Polymorphism” refers to the sequence variation observed in anindividual at a polymorphic site. Polymorphisms include nucleotidesubstitutions, insertions, deletions and microsatellites and may, butneed not, result in detectable differences in gene expression or proteinfunction. In the absence of evidence of an effect on expression orprotein function, common polymorphisms, including non-synonymousvariants, are generally considered to be included in the definition ofwild-type gene sequence. A catalog of human polymorphisms and associatedannotation, including validation, observed frequencies, and diseaseassociation, is maintained by NCBI (dbSNP: http://www.ncbi.nlmnih.gov/projects/SNP/). Please note that the term “polymorphism” whenused in the context of gene sequences should not be confused with theterm “polymorphism” when used in the context of solid state form of acompound that is the crystalline or amorphous nature of a compound. Theskilled person will understand the intended meaning by its context.

“Probe” refers to single stranded sequence-specific oligonucleotideswhich have a sequence that is exactly complementary to the targetsequence of the allele to be detected.

“Response” is defined by measurements taken according to ResponseEvaluation Criteria in Solid Tumours (RECIST) involving theclassification of patients into two main groups: those that show apartial response or stable disease and those that show signs ofprogressive disease.

“Stringent hybridisation conditions” refers to an overnight incubationat 42° C. in a solution comprising 50% formamide, 5×SSC (750 mM NaCl, 75mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5×Denhardt'ssolution, 10% dextran sulphate, and 20 pg/mI denatured, sheared salmonsperm DNA, followed by washing the filters in 0.1×SSC at about 65° C.

“Survival” encompasses a patients' overall survival and progression-freesurvival.

“Overall survival” (OS) is defined as the time from the initiation ofdrug administration to death from any cause. “Progression-free survival”(PFS) is defined as the time from the initiation of drug administrationto first appearance of progressive disease or death from any cause.

According to one aspect of the specification there is provided a methodfor selecting a patient for treatment with a compound of Formula (I),(IA), (IB), (IC) or (ID), the method comprising providing a tumour cellcontaining sample from a patient;

determining whether the ERα gene in the patient's tumour cell containingsample is wild type or mutant; and selecting a patient for treatmentwith a compound of Formula (I), (IA), (IB), (IC) or (ID) based thereon.

The method may include or exclude the actual patient sample isolationstep. Thus, according to one aspect of the specification there isprovided a method for selecting a patient for treatment with a compoundof Formula (I), (IA), (IB), (IC) or (ID), the method comprisingdetermining whether the ERα gene in a tumour cell containing samplepreviously isolated from the patient is wild type or mutant; andselecting a patient for treatment with a compound of Formula (I), (IA),(IB), (IC) or (ID) based thereon.

In one embodiment, the patient is selected for treatment with a compoundof Formula (I) if the tumour cell DNA has a mutant ERα gene. In otherembodiments, a patient whose tumour cell DNA possesses a wild type ERαgene is selected for treatment with a compound of Formula (I), (IA),(IB), (IC) or (ID).

For the purpose of this specification, a gene status of wild-type ismeant to indicate normal or appropriate expression of the gene andnormal function of the encoded protein.

In contrast, mutant status is meant to indicate expression of a proteinwith altered function, consistent with the known roles of mutant ERαgenes in cancer (as described herein). Any number of genetic orepigenetic alterations, including but not limited to mutation,amplification, deletion, genomic rearrangement, or changes inmethylation profile, may result in a mutant status. However, if suchalterations nevertheless result in appropriate expression of the normalprotein, or a functionally equivalent variant, then the gene status isregarded as wild-type. Examples of variants that typically would notresult in a functional mutant gene status include synonymous codingvariants and common polymorphisms (synonymous or non-synonymous). Asdiscussed below, gene status can be assessed by a functional assay, orit may be inferred from the nature of detected deviations from areference sequence.

In certain embodiments the wild-type or mutant status of the ERα gene isdetermined by the presence or absence of non-synonymous nucleic acidvariations in the genes. Observed non-synonymous variationscorresponding to known common polymorphisms with no annotated functionaleffects do not contribute to a gene status of mutant.

Other variations in the ERα gene that signify mutant status includesplice site variations that decrease recognition of an intron/exonjunction during processing of pre-mRNA to mRNA. This can result in exonskipping or the inclusion of normally intronic sequence in spliced mRNA(intron retention or utilization of cryptic splice junctions). This can,in turn, result in the production of aberrant protein with insertionsand/or deletions relative to the normal protein. Thus, in otherembodiments, the gene has a mutant status if there is a variant thatalters splice site recognition sequence at an intron/exon junction.

For ESR1, reference sequences are available for the gene (GenBankaccession number: NG_008493), mRNA (GenBank accession number:NM_000125), and protein (GenBank accession number: NP_000116 orSwiss-Prot accession: P03372). A person of skill in the art will be ableto determine the ESR1 gene status, i.e. whether a particular ESR1gene iswild type or mutant, based on comparison of DNA or protein sequence withwild type.

It will be apparent that the gene and mRNA sequences disclosed for ERαgene are representative sequences. In normal individuals there are twocopies of each gene, a maternal and paternal copy, which will likelyhave some sequence differences, moreover within a population there willexist numerous allelic variants of the gene sequence. Other sequencesregarded as wild type include those that possess one or more synonymouschanges to the nucleic acid sequence (which changes do not alter theencoded protein sequence), non-synonymous common polymorphisms (e.g.germ-line polymorphisms) which alter the protein sequence but do notaffect protein function, and intronic non-splice-site sequence changes.

There are numerous techniques available to the person skilled in the artto determine the gene status of ERα. The gene status can be determinedby determination of the nucleic acid sequence. This could be via directsequencing of the full-length gene or analysis of specific sites withinthe gene, e.g. commonly mutated sites.

Samples

The patient's sample to be tested for the gene status can be any tumourtissue or tumour-cell containing sample obtained or obtainable from theindividual. The test sample is conveniently a sample of blood, mouthswab, biopsy, or other body fluid or tissue obtained from an individual.Particular examples include: circulating tumour cells, circulating DNAin the plasma or serum, cells isolated from the ascites fluid of ovariancancer patients, lung sputum for patients with tumours within the lung,a fine needle aspirate from a breast cancer patient, urine, peripheralblood, a cell scraping, a hair follicle, a skin punch or a buccalsample.

It will be appreciated that the test sample may equally be a nucleicacid sequence corresponding to the sequence in the test sample, that isto say that all or a part of the region in the sample nucleic acid mayfirstly be amplified using any convenient technique e.g. polymerasechain reaction (PCR), before analysis. The nucleic acid may be genomicDNA or fractionated or whole cell RNA. In particular embodiments the RNAis whole cell RNA and is used directly as the template for labelling afirst strand cDNA using random primers or poly A primers. The nucleicacid or protein in the test sample may be extracted from the sampleaccording to standard methodologies (see Green & Sambrook, Eds.,Molecular Cloning: A Laboratory Manual, (2012, 4th edition, Vol. 1-3,ISBN 9781936113422), Cold Spring Harbor Laboratory Press, Cold SpringHarbor, N.Y.) The diagnostic methods of the specification can beundertaken using a sample previously taken from the individual orpatient. Such samples may be preserved by freezing or fixed and embeddedin formalin-paraffin or other media. Alternatively, a fresh tumour cellcontaining sample may be obtained and used.

The methods of the specification can be applied using cells from anytumour. Suitable tumours for treatment with a compound of Formula (I)have been described hereinbefore.

Methods for Detection of Nucleic Acids

The detection of mutant ERα nucleic acids can be employed, in thecontext of the present specification, to select drug treatment. Sincemutations in these genes occur at the DNA level, the methods of thespecification can be based on detection of mutations or variances ingenomic DNA, as well as transcripts and proteins themselves. It can bedesirable to confirm mutations in genomic DNA by analysis of transcriptsand/or polypeptides, in order to ensure that the detected mutation isindeed expressed in the subject.

It will be apparent to the person skilled in the art that there are alarge number of analytical procedures which may be used to detect thepresence or absence of variant nucleotides at one or more positions in agene. In general, the detection of allelic variation requires a mutationdiscrimination technique, optionally an amplification reaction (such asone based on polymerase chain reaction) and optionally a signalgeneration system. There are a multitude of mutation detectiontechniques available in the art and these may be used in combinationwith a signal generation system, of which there are numerous availablein the art. Many methods for the detection of allelic variation arereviewed by Nollau et al., Clin. Chem., 1997, 43, 1114-1120; Anderson SM. Expert Rev Mol Diagn., 2011, 11, 635-642; Meyerson M. et al., Nat RevGenet., 2010, 11, 685-696; and in standard textbooks, for example“Laboratory Protocols for Mutation Detection”, Ed. by U. Landegren,Oxford University Press, 1996 and “PCR”, 2^(nd) Edition by Newton &Graham, BIOS Scientific Publishers Limited, 1997.

As noted above, determining the presence or absence of a particularvariance or plurality of variances in the ERα gene in a patient withcancer can be performed in a variety of ways. Such tests are commonlyperformed using DNA or RNA collected from biological samples, e.g.,tissue biopsies, urine, stool, sputum, blood, cells, tissue scrapings,breast aspirates or other cellular materials, and can be performed by avariety of methods including, but not limited to, PCR, hybridizationwith allele-specific probes, enzymatic mutation detection, chemicalcleavage of mismatches, mass spectrometry or DNA sequencing, includingminisequencing.

Suitable mutation detection techniques include amplification refractorymutation system (ARMS™), amplification refractory mutation system linearextension (ALEX™), competitive oligonucleotide priming system (COPS),Taqman, Molecular Beacons, restriction fragment length polymorphism(RFLP), and restriction site based PCR and fluorescence resonance energytransfer (FRET) techniques.

In particular embodiments the method employed for determining thenucleotide(s) within a biomarker gene is selected from: allele-specificamplification (allele specific PCR)—such as amplification refractorymutation system (ARMS), sequencing, allelic discrimination assay,hybridisation, restriction fragment length polymorphism (RFLP) oroligonucleotide ligation assay (OLA).

In particular embodiments, hybridization with allele specific probes canbe conducted by: (1) allele specific oligonucleotides bound to a solidphase (e.g. glass, silicon, nylon membranes) with the labelled sample insolution, for example as in many DNA chip applications; or, (2) boundsample (often cloned DNA or PCR amplified DNA) and labelledoligonucleotides in solution (either allele specific or short so as toallow sequencing by hybridization). Diagnostic tests may involve a panelof variances, often on a solid support, which enables the simultaneousdetermination of more than one variance. Such hybridization probes arewell known in the art (see, e.g., Green & Sambrook, Eds., MolecularCloning: A Laboratory Manual, (2012, 4th edition, Vol. 1-3, ISBN9781936113422), Cold Spring Harbor Laboratory Press, Cold Spring Harbor,N.Y.) and may span two or more variance sites.

Thus, in one embodiment, the detection of the presence or absence of atleast one mutation provides for contacting ERα nucleic acid containing aputative mutation site with at least one nucleic acid probe. The probepreferentially hybridizes with a nucleic acid sequence including avariance site and containing complementary nucleotide bases at thevariance site under selective hybridization conditions. Hybridizationcan be detected with a detectable label using labels known to oneskilled in the art. Such labels include, but are not limited toradioactive, fluorescent, dye, and enzymatic labels.

In another embodiment, the detection of the presence or absence of atleast one mutation provides for contacting ERα nucleic acid containing aputative mutation site with at least one nucleic acid primer. The primerpreferentially hybridizes with a nucleic acid sequence including avariance site and containing complementary nucleotide bases at thevariance site under selective hybridization conditions.

Oligonucleotides used as primers for specific amplification may carrythe complementary nucleotide base to the mutation of interest in thecentre of the molecule (so that amplification depends on differentialhybridization; see, e.g., Gibbs, et al., 1989. Nucl. Acids Res., 17,2437-248) or at the extreme 3′-terminus of one primer where, underappropriate conditions, mismatch can prevent, or reduce polymeraseextension (see, e.g., Prossner, 1993, Tibtech, 11 238).

In yet another embodiment, the detection of the presence or absence ofat least one mutation comprises sequencing at least one nucleic acidsequence and comparing the obtained sequence with the known wild typenucleic acid sequence.

Alternatively, the presence or absence of at least one mutationcomprises mass spectrometric determination of at least one nucleic acidsequence.

In one embodiment, the detection of the presence or absence of at leastone nucleic acid variance comprises performing a polymerase chainreaction (PCR). The target nucleic acid sequence containing thehypothetical variance is amplified and the nucleotide sequence of theamplified nucleic acid is determined. Determining the nucleotidesequence of the amplified nucleic acid comprises sequencing at least onenucleic acid segment. Alternatively, amplification products can beanalysed using any method capable of separating the amplificationproducts according to their size, including automated and manual gelelectrophoresis, and the like.

Mutations in genomic nucleic acid are advantageously detected bytechniques based on mobility shift in amplified nucleic acid fragments.For instance, Chen et al., Anal Biochem 1996, 239, 61-9, describe thedetection of single-base mutations by a competitive mobility shiftassay. Moreover, assays based on the technique of Marcelino et al.,BioTechniques 1999, 26, 1134-1148 are available commercially.

In a particular example, capillary heteroduplex analysis may be used todetect the presence of mutations based on mobility shift of duplexnucleic acids in capillary systems as a result of the presence ofmismatches.

Generation of nucleic acids for analysis from samples generally requiresnucleic acid amplification. Many amplification methods rely on anenzymatic chain reaction (such as a polymerase chain reaction, a ligasechain reaction, or a self-sustained sequence replication) or from thereplication of all or part of the vector into which it has been cloned.Preferably, the amplification according to the specification is anexponential amplification, as exhibited by for example the polymerasechain reaction.

Many target and signal amplification methods have been described in theliterature, for example, general reviews of these methods in Landegren,U., et al., Science, 1988 242, 229-237 and Lewis, R., GeneticEngineering News 1990, 10, 54-55. These amplification methods can beused in the methods of our specification, and include polymerase chainreaction (PCR), PCR in situ, ligase amplification reaction (LAR), ligasehybridisation, Qβ bacteriophage replicase, transcription-basedamplification system (TAS), genomic amplification with transcriptsequencing (GAWTS), nucleic acid sequence-based amplification (NASBA)and in situ hybridisation. Primers suitable for use in variousamplification techniques can be prepared according to methods known inthe art.

Polymerase Chain Reaction (PCR) PCR is a nucleic acid amplificationmethod described inter alia in U.S. Pat. Nos. 4,683,195 and 4,683,202.PCR consists of repeated cycles of DNA polymerase generated primerextension reactions. The target DNA is heat denatured and twooligonucleotides, which bracket the target sequence on opposite strandsof the DNA to be amplified, are hybridised. These oligonucleotidesbecome primers for use with DNA polymerase. The DNA is copied by primerextension to make a second copy of both strands. By repeating the cycleof heat denaturation, primer hybridisation and extension, the target DNAcan be amplified a million fold or more in about two to four hours. PCRis a molecular biology tool, which must be used in conjunction with adetection technique to determine the results of amplification. Anadvantage of PCR is that it increases sensitivity by amplifying theamount of target DNA by 1 million to 1 billion fold in approximately 4hours. PCR can be used to amplify any known nucleic acid in a diagnosticcontext (Mok et al., Gynaecologic Oncology, 1994, 52: 247-252,).

An allele specific amplification technique such as AmplificationRefractory Mutation System (ARMS™) (Newton et al., Nucleic Acids Res.,1989, 17, 2503-2516) can also be used to detect single base mutations.Under the appropriate PCR amplification conditions a single basemismatch located at the 3′-end of the primer is sufficient forpreferential amplification of the perfectly matched allele (Newton etal., 1989, supra), allowing the discrimination of closely relatedspecies. The basis of an amplification system using the primersdescribed above is that oligonucleotides with a mismatched 3′-residuewill not function as primers in the PCR under appropriate conditions.This amplification system allows genotyping solely by inspection ofreaction mixtures after agarose gel electrophoresis.

Analysis of amplification products can be performed using any methodcapable of separating the amplification products according to theirsize, including automated and manual gel electrophoresis, massspectrometry, and the like.

The methods of nucleic acid isolation, amplification and analysis areroutine for one skilled in the art and examples of protocols can befound, for example, Green & Sambrook, Eds., Molecular Cloning: ALaboratory Manual, (2012, 4th edition, Vol. 1-3, ISBN 9781936113422),Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.)Particularly useful protocol source for methods used in PCRamplification is PCR (Basics: From Background to Bench) by M. J.McPherson, S. G. Mailer, R. Beynon, C. Howe, Springer Verlag; 1stedition (Oct. 15, 2000), ISBN: 0387916008.

The present specification also provides predictive and diagnostic kitscomprising degenerate primers to amplify a target nucleic acid in theERα gene and instructions comprising; amplification protocol andanalysis of the results. The kit may alternatively also comprisebuffers, enzymes, and containers for performing the amplification andanalysis of the amplification products. The kit may also be a componentof a screening, or diagnostic kit comprising other tools such as DNAmicroarrays, or other supports. Preferably, the kit also provides one ormore control templates, such as nucleic acids isolated from normaltissue sample, and/or a series of samples representing differentvariances in the reference genes.

In one embodiment, the kit provides two or more primer pairs, each paircapable of amplifying a different region of the reference (ERα) gene(each region a site of potential variance) thereby providing a kit foranalysis of expression of several gene variances in a biological samplein one reaction or several parallel reactions.

Primers in the kits may be labelled, for example fluorescently labelled,to facilitate detection of the amplification products and consequentanalysis of the nucleic acid variances. The kit may also allow for morethan one variance to be detected in one analysis. A combination kit willtherefore comprise of primers capable of amplifying different segmentsof the reference gene. The primers may be differentially labelled, forexample using different fluorescent labels, so as to differentiatebetween the variances.

In another aspect, the specification provides a method of treating apatient suffering from cancer comprising: determining the mutant or wildtype status of the ERα gene in the patient's tumour cells and if the ERαgene is mutant, administering to the patient an effective amount of acompound of Formula (I), (IA), (IB), (IC) or (ID).

As used herein, the terms “effective” and “effectiveness” includes bothpharmacological effectiveness and physiological safety. Pharmacologicaleffectiveness refers to the ability of the treatment to result in adesired biological effect in the patient.

Physiological safety refers to the level of toxicity, or other adversephysiological effects at the cellular, organ and/or organism level(often referred to as side-effects) resulting from administration of thetreatment. “Less effective” means that the treatment results in atherapeutically significant lower level of pharmacological effectivenessand/or a therapeutically greater level of adverse physiological effects.

According to another aspect of the specification there is provided theuse of a compound of Formula (I), (IA), (IB), (IC) or (ID), or apharmaceutically acceptable salt thereof to treat a cancer patient whosetumour cells have been identified as possessing a mutant ERα gene.

According to another aspect of the specification there is provided acompound of Formula (I), (IA), (IB), (IC) or (ID), or a pharmaceuticallyacceptable salt thereof for treating cancers with tumour cellsidentified as harbouring mutant ERα gene.

According to another aspect of the specification there is provided amethod of treating cancers with tumour cells identified as harbouringmutant ERα gene comprising administering an effective amount of acompound of Formula (I), (IA), (IB), (IC) or (ID), or a pharmaceuticallyacceptable salt thereof.

In still further embodiments, the specification relates to apharmaceutical composition comprising a compound of Formula (I), (IA),(IB), (IC) or (ID), for use in the prevention and treatment of cancerwith tumour cells identified as harbouring a mutant ERα gene.

For all the aspects above, mutant forms of ERα determined/identified areat all positions across the gene.

For all the aspects above, using tumours such as breast cancer as anexample, particular mutant forms of ERα determined/identified are thoseat positions Ser463Pro, Va1543Glu, Leu536Arg, Tyr537Ser, Tyr537Asn andAsp538Gly.

DESCRIPTION OF DRAWINGS

FIG. 1 shows the X-Ray Powder Diffraction Pattern for Form A ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine

FIG. 2 shows the DSC/TGA Thermogram for Form A ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine.

FIG. 3 shows the X-Ray Powder Diffraction Pattern for Form B ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine.

FIG. 4 shows the X-Ray Powder Diffraction Pattern for Form C ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine.

FIG. 5 shows the DSC/TGA Thermogram for Form C ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine.

FIG. 6 shows the X-Ray Powder Diffraction Pattern for Form D ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine.

FIG. 7 shows the DSC/TGA Thermogram for Form D ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine.

FIG. 8 shows the X-Ray Powder Diffraction Pattern for Form E ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine

FIG. 9 shows the DSC/TGA Thermogram for Form E ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine.

FIG. 10 shows the X-Ray Powder Diffraction Pattern for Form F ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine.

FIG. 11 shows the X-Ray Powder Diffraction Pattern for Form G ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine.

FIG. 12 shows the results of a human parental MCF7 xenograft anti-tumourefficacy study in mouse withN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine.

FIG. 13 shows the results of a human Y537S ESR1 mutant MCF7 xenograftanti-tumour efficacy study in mouse withN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine.

FIG. 14 shows the results of a human ESR1 mutant breast cancer patientderived xenograft CTC174 anti-tumour efficacy study in mouse withN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine.

FIG. 15 shows the results of a human ESR1 mutant breast cancer patientderived xenograft CTC174 anti-tumour efficacy study in mouse with acombination ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amineand palbociclib.

FIG. 16 shows the results of a human ESR1 mutant breast cancer patientderived xenograft CTC174 anti-tumour efficacy study in mouse with acombination ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amineand vistusertib (AZD2014).

EXAMPLES

The compounds described in this specification are further illustrated inthe following Examples. These Examples are given by way of illustrationonly and are non-limiting. In general:

(i) operations were carried out at ambient temperature, i.e. in therange 17 to 25° C. and under an atmosphere of an inert gas such asnitrogen unless otherwise stated;

(ii) evaporations were carried out by rotary evaporation or utilisingGenevac equipment or Biotage v10 evaporator in vacuo and work-upprocedures were carried out after removal of residual solids byfiltration;

(iii) flash chromatography purifications were performed on an automatedTeledyne Isco CombiFlash® Rf or Teledyne Isco CombiFlash® Companion®using prepacked RediSep Rf Gold™ Silica Columns (20-40 μm, sphericalparticles), GraceResolv™ Cartridges (Davisil® silica) or Silicyclecartridges (40-63 μm).

(iv) preparative chromatography was performed on a Gilson prep HPLCinstrument with UV collection or via supercritical fluid chromatographyperformed on a Waters Prep 100 SFC-MS instrument with MS- andUV-triggered collection or a Thar MultiGram III SFC instrument with UVcollection;

(v) chiral preparative chromatography was performed on a Gilsoninstrument with UV collection (233 injector/fraction collector, 333 &334 pumps, 155 UV detector) or a Varian Prep Star instrument (2×SD1pumps, 325 UV detector, 701 fraction collector) pump running with Gilson305 injection;

(vi) yields, where present, are not necessarily the maximum attainable;

(vii) in general, the structures of end-products of the Formula I wereconfirmed by nuclear magnetic resonance (NMR) spectroscopy; NMR chemicalshift values were measured on the delta scale [proton magnetic resonancespectra were determined using a Bruker Avance 500 (500 MHz) or BrukerAvance 400 (400 MHz) instrument]; measurements were taken at ambienttemperature unless otherwise specified; the following abbreviations havebeen used: s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet;dd, doublet of doublets; ddd, doublet of doublet of doublet; dt, doubletof triplets; bs, broad signal

(viii) in general, end-products of the Formula I were also characterisedby mass spectroscopy following liquid chromatography (LCMS or UPLC);UPLC was carried out using a Waters UPLC fitted with Waters SQ massspectrometer (Column temp 40, UV=220-300 nm, Mass Spec=ESI withpositive/negative switching) at a flow rate of 1 ml/min using a solventsystem of 97% A+3% B to 3% A to 97% B over 1.50 mins (total runtime withequilibration back to starting conditions etc 1.70 min), where A=0.1%formic acid in water (for acid work) or 0.1% ammonia in water (for basework) B=acetonitrile. For acid analysis the column used was WatersAcquity HSS T3 1.8 μm 2.1×50 mm, for base analysis the column used wasWaters Acquity BEH 1.7 μm 2.1×50 mm; LCMS was carried out using a WatersAlliance HT (2795) fitted with a Waters ZQ ESCi mass spectrometer and aPhenomenex Gemini-NX (50×2.1 mm 5 μm) column at a flow rate of 1.1ml/min 95% A to 95% B over 4 min with a 0.5 min hold. The modifier iskept at a constant 5% C (50:50 acetonitrile:water 0.1% formic acid) or D(50:50 acetonitrile:water 0.1% ammonium hydroxide (0.88 SG) depending onwhether it is an acidic or basic method.

(ix) ion exchange purification was generally performed using a SCX-2(Biotage, Propylsulfonic acid functionalized silica. Manufactured usinga trifunctional silane. Non end-capped) cartridge.

(x) intermediate purity was assessed by thin layer chromatographic, massspectral, HPLC (high performance liquid chromatography) and/or NMRanalysis;

(xi) the following abbreviations have been used:—

-   -   AcOH acetic acid    -   aq. Aqueous    -   Boc tert-butoxycarbonyl    -   n-BuLi n-butyl lithium    -   tBuOH tert-butanol    -   Brettphos        2-(dicyclohexylphosphino)3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl    -   CDCl₃ deutero-chloroform

CHCl₃ chloroform

-   -   Conc. concentrated    -   DCM dichloromethane    -   DIPEA diisopropylethylamine    -   DMAP dimethylaminopyridine    -   DMSO dimethyl sulphoxide    -   EtOH ethanol    -   EtOAc ethyl acetate    -   HATU        1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium        3-oxid hexafluorophosphate    -   HCl hydrochloric acid    -   HPLC high performance liquid chromatography    -   K₂CO₃ potassium carbonate    -   MeOH methanol    -   MgSO₄ magnesium sulfate    -   NaH sodium hydride    -   NaHCO₃ sodium bicarbonate    -   Na₂SO₄ sodium sulfate    -   Pd₂(dba)₃ tris(dibenzylideneacetone)dipalladium(0)    -   Rac-BINAP (±)-2,2′-Bis(diphenylphosphino)-1,1′-binaphthalene    -   rt/RT room temperature    -   RockPhos 3^(rd) generation        [(2-Di-tert-butylphosphino-3-methoxy-6-precatalyst        methyl-2′,4′,6′-triisopropyl-1,1′-biphenyl)-2-(2-aminobiphenyl)]palladium(II)        methanesulfonate    -   Ruphos 2-Dicyclohexylphosphino-2′,6′-diisopropoxy-1,1′-biphenyl    -   sat. saturated    -   SFC supercritical fluid chromatography    -   sol. solution    -   TBAF tetra-n-butylammonium fluoride    -   TEA triethylamine    -   THF tetrahydrofuran    -   THP tetrahydropyran    -   TFA trifluoroacetic acid    -   Xantphos 4,5-bis(diphenylphosphino)-9,9-dimethyl xanthene

Example 1N-(4-((6S,8R)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine

[(2-Di-cyclohexylphosphino-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl)-2-(2′-amino-1,1′-biphenyl)]palladium(II)methanesulfonate (BrettPhos Pd G3) (6.80 mg, 7.50 μmol) was added to asuspension of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(85 mg, 0.15 mmol), 1-(3-fluoropropyl)azetidin-3-amine (29.7 mg, 0.23mmol) and sodium tert-butoxide (28.8 mg, 0.30 mmol) in degassed1,4-dioxane (1.5 mL) and the reaction was heated to 90° C. for 3 hours.After cooling, the reaction was diluted with DCM and washed with water.The aqueous was extracted with DCM, then the combined organics weredried and evaporated. The crude residue was dissolved in DCM (2 mL) andTFA (0.7 mL) was added. The reaction was stirred at room temperature for1 hour, then it was diluted with DCM and basified by addition ofsaturated NaHCO₃ solution. The layers were separated and the aqueous wasextracted with DCM. The combined organics were dried and evaporated,then the crude product was purified by flash silica chromatography,elution gradient 0 to 20% MeOH in EtOAc. Pure fractions were evaporatedto dryness to affordN-(4-((6S,8R)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine(39.0 mg, 49%) as a beige solid. ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.09(3H, d), 1.83 (2H, ddd), 2.65-2.86 (4H, m), 2.96-3.08 (1H, m), 3.11 (1H,dd), 3.22 (2H, d), 3.38 (3H, s), 3.51-3.63 (2H, m), 3.73-3.82 (1H, m),3.82 (3H, s), 3.86 (2H, q), 4.15-4.28 (1H, m), 4.43 (1H, t), 4.53 (1H,t), 5.37 (1H, s), 5.88 (1H, dd), 6.14 (1H, d), 6.46 (1H, d), 6.80 (1H,d), 7.13 (1H, d), 8.04 (1H, d). m/z: ES+ [M+H]+ 532.

The(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolinewas prepared as follows:

Preparation of 2,2-difluoro-3-(trityloxy)propan-1-ol

2,2-Difluoropropane-1,3-diol (2.50 g, 22.3 mmol) was dissolved in DCM(61.7 mL) and THF (15.4 mL). DIPEA (3.93 mL, 22.3 mmol) was added,followed by (chloromethanetriyl)tribenzene (6.22 g, 22.3 mmol) andfinally DMAP (0.288 g, 2.23 mmol). The reaction was heated to 40° C. for2 hours. After cooling, the reaction was washed with 1N HCl solution,then dried and evaporated. The crude product was purified by flashsilica chromatography, elution gradient 0 to 25% EtOAc in heptane. Purefractions were evaporated to dryness to afford2,2-difluoro-3-(trityloxy)propan-1-ol (4.43 g, 56%) as a colourlesssolid. ¹H NMR (500 MHz, CDCl₃) 3.42 (2H, t), 3.92 (2H, t), 7.23-7.3 (4H,m), 7.3-7.39 (6H, m), 7.39-7.49 (6H, m).

Preparation of ((2,2-difluoro-3-methoxypropoxy)methanetriyl)tribenzene

Sodium hydride (0.562 g, 14.0 mmol) was added to a solution of2,2-difluoro-3-(trityloxy)propan-1-ol (4.15 g, 11.7 mmol) in THF (46 mL)and the reaction was stirred for 1 hour, then iodomethane (0.802 mL,12.9 mmol) was added in THF (5 mL). The reaction was stirred for afurther 1 hour. The reaction was quenched with water and brine, thenextracted with EtOAc. The organic layer was dried and evaporated toafford ((2,2-difluoro-3-methoxypropoxy)methanetriyl)tribenzene (4.18 g,97%) as a pale yellow oil. ¹H NMR (500 MHz, CDCl₃) 3.36 (2H, t), 3.40(3H, s), 3.77 (2H, t), 7.15-7.28 (3H, m), 7.28-7.38 (6H, m), 7.39-7.47(6H, m).

Preparation of 2,2-difluoro-3-methoxypropyl trifluoromethanesulfonate

Trifluoromethanesulfonic anhydride (1.918 mL, 11.40 mmol) was added to asolution of ((2,2-difluoro-3-methoxypropoxy)methanetriyl)tribenzene(4.00 g, 10.9 mmol) in DCM (39.6 mL). The reaction was stirred for 30minutes, then triethylsilane (1.934 mL, 11.94 mmol) was added and thereaction was stirred for a further 30 minutes. The reaction wasevaporated, and the triflate, was used directly in the next stage.

Preparation of (R)-1-(3-bromo-2-methylphenyl)propan-2-amine

n-BuLi (26.1 mL, 41.8 mmol) was added dropwise to a solution of1,3-dibromo-2-methylbenzene (9.95 g, 39.8 mmol) in THF (100 mL) at −78°C. After stirring for 30 minutes, (R)-tert-butyl4-methyl-1,2,3-oxathiazolidine-3-carboxylate 2,2-dioxide (10.39 g, 43.8mmol) was added in portions and the reaction was stirred for a further30 minutes before being allowed to warm to 0° C. over 30 minutes. 1Ncitric acid was added and the mixture was stirred for 5 minutes beforeit was extracted with EtOAc (×2). The combined organic phases wereevaporated. The residue was stirred in 4M HCl in dioxane (69.7 mL, 278.7mmol) at room temperature for 1 hour, then the volatiles wereevaporated. The residue was suspended in diethyl ether, and extractedwith water (×2). The combined aqueous phases were basified by additionof 2N Na₂CO₃, then extracted with DCM (×3). The combined organics phaseswere dried over MgSO₄ and concentrated to afford(R)-1-(3-bromo-2-methylphenyl)propan-2-amine (7.55 g, 83%) as a yellowoil. ¹H NMR (400 MHz, CDCl₃, 27° C.) 1.13 (3H, d), 1.43 (2H, s), 2.40(3H, s), 2.61 (1H, dd), 2.77 (1H, dd), 3.14 (1H, dq), 6.97 (1H, t), 7.08(1H, d), 7.43 (1H, d). m/z (ES+), [M+H]+=228.

Preparation of(R)—N-(1-(3-bromo-2-methylphenyl)propan-2-yl)-2,2-difluoro-3-methoxypropan-1-amine

2,2-Difluoro-3-methoxypropyl trifluoromethanesulfonate (3.03 g, 11.73mmol) (crude from the previous step) was added to a solution of(R)-1-(3-bromo-2-methylphenyl)propan-2-amine (2.327 g, 10.2 mmol) andDIPEA (2.82 mL, 16.32 mmol) in 1,4-dioxane (34.3 mL) and the reactionwas heated to 80° C. overnight. After cooling, the volatiles wereevaporated, then the residue was dissolved in DCM and washed with brine.The organic phase was dried and evaporated, then the crude product waspurified by flash silica chromatography, elution gradient 0 to 100%EtOAc in heptane. Pure fractions were evaporated to dryness to afford(R)—N-(1-(3-bromo-2-methylphenyl)propan-2-yl)-2,2-difluoro-3-methoxypropan-1-amine(2.330 g, 68%) as a pale yellow oil. ¹H NMR (500 MHz, CDCl₃) 1.06 (3H,d), 2.40 (3H, s), 2.63 (1H, dd), 2.82 (1H, dd), 2.86-2.94 (1H, m),2.95-3.11 (2H, m), 3.38 (3H, s), 3.51-3.63 (2H, m), 6.94-7 (1H, m), 7.07(1H, dd), 7.4-7.51 (1H, m). m/z: ES+ [M+H]+ 336.

Preparation of(R)-3-(24(2,2-difluoro-3-methoxypropyl)amino)propyl)-2-methylaniline

Pd₂(dba)₃ (0.184 g, 0.20 mmol) and Rac-BINAP (0.250 g, 0.40 mmol) wereadded to a suspension of(R)—N-(1-(3-bromo-2-methylphenyl)propan-2-yl)-2,2-difluoro-3-methoxypropan-1-amine(2.25 g, 6.69 mmol), benzophenone imine (1.334 g, 7.36 mmol) and sodiumtert-butoxide (0.965 g, 10.0 mmol) in degassed toluene (28.5 mL) and thereaction was heated to 90° C. for 3 hours. After cooling, the toluenewas largely evaporated, then the residue was dissolved in DCM and washedwith water. The aqueous phase was extracted with DCM, then the organicswere evaporated to ˜50 mL volume. 2N HCl solution (50 mL) was added andthe biphasic mixture was stirred vigorously for 30 minutes. The layerswere separated, then the aqueous was extracted with DCM. The organicphase was back extracted with 1N HCl. The combined aqueous phases werebasified by addition of solid K₂CO₃ then extracted with DCM (×3), andthe combined DCM extracts were dried and evaporated to afford(R)-3-(2-((2,2-difluoro-3-methoxypropyl)amino)propyl)-2-methylaniline(1.780 g, 98%) as a light brown oil. ¹H NMR (500 MHz, CDCl₃) 1.06 (3H,d), 2.11 (3H, s), 2.59 (1H, dd), 2.75 (1H, dd), 2.86-2.94 (1H, m),2.94-3.12 (2H, m), 3.38 (3H, s), 3.53-3.63 (4H, m), 6.5-6.68 (2H, m),6.86-7.01 (1H, m). m/z: ES+ [M+H]+ 273.

Preparation of(1S,3R)-1-(4-bromo-2-methoxyphenyl)-2-(2,2-difluoro-3-methoxypropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine

(R)-3-(2-((2,2-Difluoro-3-methoxypropyl)amino)propyl)-2-methylaniline(490 mg, 1.80 mmol) and 4-bromo-2-methoxybenzaldehyde (813 mg, 3.78mmol) were heated in acetic acid (8.8 mL) and water (0.162 mL, 9.00mmol) to 75° C. overnight. After cooling, the acetic acid was evaporatedunder vacuum, then the residue was dissolved in EtOAc (20 mL) and 2N HClsolution (20 mL) was added. The biphasic mixture was stirred for 30minutes, then the layers were separated. The organic phase was extractedwith water, then the aqueous phase was basified by addition of 2N NaOHsolution and extracted with DCM (×2). The combined DCM layers were driedand evaporated, then the crude product was purified by flash silicachromatography, elution gradient 0 to 50% EtOAc in heptane. Purefractions were evaporated to dryness to afford(1S,3R)-1-(4-bromo-2-methoxyphenyl)-2-(2,2-difluoro-3-methoxypropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(384 mg, 46%) as a beige solid. ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.03(3H, d), 2.07 (3H, s), 2.42 (1H, dd), 2.60-2.76 (2H, m), 2.99 (1H, ddd),3.34 (1H, d), 3.36 (3H, s), 3.52 (2H, s), 3.54-3.66 (1H, m), 3.76 (1H,ddd), 3.87 (3H, s), 5.28 (1H, s), 6.47 (2H, s), 6.59 (1H, d), 6.88 (1H,dd), 7.01 (1H, d). m/z: ES+ [M+H]+ 469.

Preparation of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

Sodium nitrite (59.8 mg, 0.87 mmol) was added in water (0.2 mL) to acooled solution of(1S,3R)-1-(4-bromo-2-methoxyphenyl)-2-(2,2-difluoro-3-methoxypropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(370 mg, 0.79 mmol) in propionic acid (2628 μL)/water (526 μL) at −10°C. The reaction was stirred for 1 hour, then ice-cold EtOAc (20 mL) wasadded. The reaction was quenched by addition of cold saturated NaHCO₃solution and stirred for 15 minutes, before being allowed to warm toroom temperature. The layers were separated and the aqueous wasextracted with EtOAc. The combined organics were dried and evaporated,then the crude product was purified by flash silica chromatography,elution gradient 0 to 50% EtOAc in heptane. Pure fractions wereevaporated to dryness to afford(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(252 mg, 67%) as a beige solid. ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.10(3H, d), 2.72 (1H, td), 2.85 (1H, dd), 3.08 (1H, ddd), 3.16 (1H, dd),3.36 (3H, s), 3.48-3.66 (2H, m), 3.65-3.81 (1H, m), 3.90 (3H, s), 5.44(1H, s), 6.59 (1H, d), 6.77 (1H, d), 6.89 (1H, dd), 7.05 (1H, d), 7.19(1H, dd), 8.07 (1H, d). m/z: ES+ [M+H]+ 480.

Preparation of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

3,4-Dihydro-2H-pyran (0.064 mL, 0.70 mmol) was added to a solution of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(240 mg, 0.50 mmol) and p-toluenesulfonic acid hydrate (9.51 mg, 0.05mmol) in DCM (2.5 mL) and the reaction was heated to 40° C. for 1 hour.After cooling, the reaction was diluted with DCM and washed withsaturated NaHCO₃ solution. The organic phase was dried and evaporated.The crude residue was passed through a plug of silica gel(EtOAc/heptane, 1:1 as eluent) and the filtrate was evaporated to afford(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(262 mg, 93%) as a pale yellow solid as a 5:1 ratio of THP regioisomers(each THP regioisomer is a diastereosiomeric mixture). m/z: ES+ [M+H]+564.

Preparation of tert-butyl (1-(3-fluoropropyl)azetidin-3-yl)carbamate

Potassium carbonate (1.605 g, 11.61 mmol), tert-butylazetidin-3-ylcarbamate (1.0 g, 5.81 mmol) and 1-fluoro-3-iodopropane(1.146 g, 6.10 mmol) were suspended in acetonitrile (11.61 mL) andsealed into a microwave tube. The reaction was heated to 95° C. for 15minutes in the microwave reactor. Reaction was cooled to roomtemperature, diluted with EtOAc (100 mL) and extracted from saturatedNaHCO₃(50 mL). The organic phase was dried over MgSO₄, filtered andevaporated to afford tert-butyl(1-(3-fluoropropyl)azetidin-3-yl)carbamate (1.248 g, 93%) as a yellowsolid. ¹H NMR (500 MHz, CDCl₃, 22° C.) 1.44 (9H, s), 1.68-1.80 (2H, m),2.56 (2H, t), 2.88 (2H, s), 3.66 (2H, t), 4.31 (1H, d), 4.43 (1H, t),4.53 (1H, t), 4.90 (1H, s).

Preparation of 1-(3-fluoropropyl)azetidin-3-amine

TFA (2.69 ml) was added to a solution of tert-butyl(1-(3-fluoropropyl)azetidin-3-yl)carbamate (1.248 g, 5.37 mmol) in DCM(8.06 mL) and the reaction was stirred at room temperature for 1 hour.The volatiles were evaporated and purified by ion exchangechromatography, using an SCX column. The desired product was eluted fromthe column using 1M NH₃/MeOH and pure fractions were evaporated todryness to afford 1-(3-fluoropropyl)azetidin-3-amine (0.697 g, 98%) as acolourless oil. ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.66-1.83 (2H, m), 1.96(3H, s), 2.54 (2H, t), 2.67 (2H, td), 3.58-3.70 (2H, m), 4.43 (1H, t),4.52 (1H, t).

Example 264(6S,8R)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine

[(2-Di-cyclohexylphosphino-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl)-2-(2′-amino-1,1′-biphenyl)]palladium(II)methanesulfonate (BrettPhos Pd G3) (6.83 mg, 8.00 μmol) and sodiumtert-butoxide (48.0 mg, 0.50 mmol) were added to a degassed solution of(6S,8R)-6-(5-bromopyridin-2-yl)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(107 mg, 0.20 mmol) and 1-(3-fluoropropyl)azetidin-3-amine (52.9 mg,0.40 mmol) in 1,4-dioxane (1.6 mL) and the reaction was heated to 90° C.for 5 hours. After cooling, the reaction was diluted with DCM and washedwith water. The organic phase was evaporated, then dissolved in DCM (2mL), before TFA (1 mL) was added. The mixture was stirred at roomtemperature for 1 hour, then was diluted with DCM and washed withsaturated NaHCO₃ solution. The layers were separated and the aqueous wasextracted with DCM. The combined organics were dried and evaporated,then the crude product was purified by flash silica chromatography,elution gradient 0 to 20% MeOH in EtOAc. Pure fractions were evaporatedto dryness to afford6-((6S,8R)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine(83 mg, 83%) as a beige solid. ¹H NMR (500 MHz, CDCl₃) 1.12 (3H, d),1.79 (2H, ddd), 2.71 (2H, t), 2.75-2.90 (2H, m), 3.06-3.23 (3H, m), 3.39(3H, s), 3.51-3.66 (2H, m), 3.66-3.76 (1H, m), 3.76-3.84 (2H, m), 4.14(1H, s), 4.44 (2H, t), 4.53 (1H, t), 5.06 (1H, s), 6.80 (1H, dd), 6.88(1H, d), 7.13 (2H, dd), 7.84 (1H, d), 7.99 (1H, d). m/z: ES+ [M+H]+ 503.

The(6S,8R)-6-(5-bromopyridin-2-yl)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolinewas prepared as follows:

Preparation of(1S,3R)-1-(5-bromopyridin-2-yl)-2-(2,2-difluoro-3-methoxypropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine

5-Bromopicolinaldehyde (1172 mg, 6.30 mmol) was added to a solution of(R)-3-(2-((2,2-difluoro-3-methoxypropyl)amino)propyl)-2-methylaniline(817 mg, 3.00 mmol) in acetic acid (14.7 mL) and water (270 μL, 15.0mmol) and the reaction was heated to 80° C. for 2 hours. After cooling,the volatiles were evaporated under vacuum. The residue was dissolved inDCM and washed with saturated NaHCO₃ solution. The organic wasevaporated to a volume ˜20 mL and 2N HCl solution (20 mL) was added. Thebiphasic mixture was stirred for 15 minutes, then separated. The organicwas extracted with water, then the aqueous phase was back-extracted withDCM. The aqueous phase was then basified by addition of solid K₂CO₃,then extracted with DCM. The organic extracts were dried and evaporated.The crude product was purified by flash silica chromatography, elutiongradient 0 to 50% EtOAc in heptane. Pure fractions were evaporated todryness to afford(1S,3R)-1-(5-bromopyridin-2-yl)-2-(2,2-difluoro-3-methoxypropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(810 mg, 61%) as a beige solid. ¹H NMR (500 MHz, CDCl₃) 1.07 (3H, d),2.05 (3H, s), 2.49 (1H, d), 2.75 (2H, dd), 3.04-3.17 (1H, m), 3.30-3.36(1H, m), 3.37 (3H, s), 3.58-3.74 (2H, m), 4.96 (1H, s), 6.51 (1H, d),6.60 (1H, d), 7.22 (1H, d), 7.68 (1H, dd), 8.55 (1H, dd). m/z: ES+[M+H]+ 440.

Preparation of(6S,8R)-6-(5-bromopyridin-2-yl)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

Sodium nitrite (133 mg, 1.93 mmol) was added in water (0.5 mL) to acooled solution of(1S,3R)-1-(5-bromopyridin-2-yl)-2-(2,2-difluoro-3-methoxypropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(771 mg, 1.75 mmol) in propionic acid (5833 μL)/water (1167 μL) at −15°C. The reaction was stirred for 30 minutes, then EtOAc (50 mL), whichhad been cooled in dry-ice was added. The reaction was quenched byaddition of 2N Na₂CO₃ until bubbling ceased, then the layers wereseparated. The aqueous was extracted with EtOAc, then the organic wasdried and evaporated. The crude product was purified by flash silicachromatography, elution gradient 0 to 50% EtOAc in heptane. Purefractions were evaporated to dryness to afford(6S,8R)-6-(5-bromopyridin-2-yl)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(460 mg, 58%) as a pale yellow solid. ¹H NMR (500 MHz, CDCl₃) 1.14 (3H,d), 2.81 (1H, dd), 2.88 (1H, dd), 3.10-3.26 (2H, m), 3.38 (3H, s),3.46-3.55 (1H, m), 3.58-3.76 (2H, m), 5.13 (1H, s), 6.94 (1H, d), 7.23(1H, dd), 7.29 (1H, d), 7.72 (1H, dd), 8.05 (1H, d), 8.57 (1H, dd). m/z:ES+ [M+H]+ 451.

Preparation of(6S,8R)-6-(5-bromopyridin-2-yl)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

3,4-Dihydro-2H-pyran (0.114 mL, 1.25 mmol) was added to a solution of(6S,8R)-6-(5-bromopyridin-2-yl)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(450 mg, 1.00 mmol) and PTSA hydrate (37.9 mg, 0.20 mmol) in DCM (5 mL)and the reaction was heated to 45° C. for 3 hours. After cooling, thereaction was diluted with DCM and washed with saturated NaHCO₃ solution.The organic phase was dried and evaporated, then the crude was passedthrough a plug of silica (1:1 EtOAc/heptane). The filtrate wasevaporated to afford(6S,8R)-6-(5-bromopyridin-2-yl)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(510 mg, 9%) as an orange solid (˜6.5:1 ratio of THP regioisomers). m/z:ES+ [M+H]+ 535.

Example 36-((6S,8R)-7-((l-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine

[(2-Di-cyclohexylphosphino-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl)-2-(2′-amino-1,1′-biphenyl)]palladium(II)methanesulfonate (BrettPhos Pd G3) (12.11 mg, 0.01 mmol) and sodiumtert-butoxide (85 mg, 0.89 mmol) were added to a degassed solution of(6S,8R)-6-(5-bromopyridin-2-yl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(177 mg, 0.35 mmol) and 1-(3-fluoropropyl)azetidin-3-amine (94 mg, 0.71mmol) in 1,4-dioxane (2835 μL) and the reaction was heated to 90° C. for5 hours. After cooling, the reaction was diluted with DCM and washedwith water. The organic phase was evaporated, then dissolved in DCM (2mL), before TFA (1 mL) was added. The mixture was stirred at roomtemperature for 1 hour, then was diluted with DCM and washed withsaturated NaHCO₃ solution. The layers were separated and the aqueousphase was extracted with DCM. The combined organics were dried andevaporated to give the crude product. The crude product was purified bypreparative LCMS (Waters SunFire column, 5μ silica, 19 mm diameter, 100mm length), using decreasingly polar mixtures of water (containing 1%NH₃) and MeCN as eluents.

Fractions containing the desired compounds were evaporated to dryness togive6-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine(17.0 mg, 10%) as a beige solid. ¹H NMR (500 MHz, CDCl₃, 27° C.)0.36-0.46 (1H, m), 0.49-0.59 (1H, m), 0.88-1.07 (2H, m), 1.09 (3H, d),1.69-1.75 (1H, m), 1.75-1.81 (1H, m), 2.59 (2H, t), 2.70 (1H, dd),2.86-2.96 (3H, m), 3.01-3.11 (1H, m), 3.37-3.44 (1H, m), 3.72 (2H, q),3.78-3.88 (1H, m), 3.95 (1H, d), 4.04-4.13 (1H, m), 4.44 (1H, t), 4.53(1H, t), 4.95 (1H, s), 6.75 (1H, dd), 6.91 (1H, d), 7.12-7.17 (2H, m),7.86 (1H, d), 8.05 (1H, d), 10.04 (1H, s). m/z: ES+ [M+H]+ 467.

The(6S,8R)-6-(5-bromopyridin-2-yl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolinewas prepared as follows:

Preparation of 1-(3-bromo-2-methylphenyl)-2,5-dimethyl-1H-pyrrole

A mixture of 3-bromo-2-methylaniline (40 g, 215 mmol), hexane-2,5-dione(25.3 mL, 215 mmol) and p-toluenesulfonic acid monohydrate (0.409 g,2.15 mmol) in toluene (300 mL) was heated at reflux conditions for 2hours in a flask equipped with a condensor and Dean-Stark trap. Themixture was then cooled to room temperature and washed sequentially withsaturated aqueous sodium bicarbonate, aqueous HCl (1N), and saturatedaqueous sodium chloride. The organic layer was dried over sodiumsulfate, filtered and concentrated under reduced pressure to give crude1-(3-bromo-2-methylphenyl)-2,5-dimethyl-1H-pyrrole (57.9 g, 102%) as apale yellow oil. ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 1.8 (6H, s), 1.9 (3H,s), 5.8 (2H, s), 7.2-7.4 (2H, m), 7.7 (1H, dd). m/z: ES+[M+H]+ 264.

Preparation of tert-butyl(R)-(1-(3-(2,5-dimethyl-1H-pyrrol-1-yl)-2-methylphenyl)propan-2-yl)carbamate

n-Butyllithium in hexane (2.5M; 89 mL, 221 mmol) was added over 15minutes to a solution of crude1-(3-bromo-2-methylphenyl)-2,5-dimethyl-1H-pyrrole (55.7 g, 211 mmol) inTHF (400 mL) at −78° C. After 30 minutes, tert-butyl(R)-4-methyl-1,2,3-oxathiazolidine-3-carboxylate 2,2-dioxide (50 g, 211mmol) was added. The resulting mixture was stirred at −78° C. for 15minutes and allowed to warm to room temperature over 2 hours. Aqueouscitric acid (1N; 250 mL) was added, and stirring was continued for 30minutes. The mixture was extracted with hexanes, and the combinedorganic layers were washed with saturated aqueous sodium carbonate,dried over sodium sulfate, filtered, and concentrated under reducedpressure. The resulting brown solid, crude tert-butyl(R)-(1-(3-(2,5-dimethyl-1H-pyrrol-1-yl)-2-methylphenyl)propan-2-yl)carbamatewas used in the next step without further purification. m/z: ES+ [M+H]+343.

Preparation of(R)-1-(3-(2,5-dimethyl-1H-pyrrol-1-yl)-2-methylphenyl)propan-2-amine

Hydrochloric acid in dioxane (4 M; 100 mL, 400 mmol) was added to asuspension of crude tert-butyl(R)-(1-(3-(2,5-dimethyl-1H-pyrrol-1-yl)-2-methylphenyl)propan-2-yl)carbamatein MeOH (200 mL) and DCM (50 mL). The resulting red solution was stirredat room temperature for 4 hours and then concentrated under reducedpressure. The resulting brown solid was used in the next step withoutfurther purification. m/z: ES+[M+H]+ 243.

Preparation of (R)-3-(2-aminopropyl)-2-methylaniline

A mixture of crude(R)-1-(3-(2,5-dimethyl-1H-pyrrol-1-yl)-2-methylphenyl)propan-2-aminedihydrochloride, aqueous hydroxylamine (50 wt %; 107 mL, 1.74 mol), andhydroxylamine hydrochloride (97 g, 1.39 mol) in ethanol (400 mL) waswarmed to reflux conditions. After 18 hours, the reaction was cooled to0° C., basified with aqueous sodium hydroxide (50 wt %; 153 g, 1.92 mol)and extracted with DCM. The organic layer was dried over sodium sulfate,filtered and concentrated under reduced pressure. The resulting residuewas purified by SFC (Princeton Chromatography DEAP column, 100 mmlength, 30 mm diameter, 5 μm, 40° C. column temperature, 100 bar columnpressure, 100 mg/mL flow rate), eluting with 25% methanol containing0.2% NH₄OH in CO₂, to afford (R)-3-(2-aminopropyl)-2-methylaniline (24g, 84%) as a light amber solid. ¹H NMR (500 MHz, DMSO, 27° C.) 1.05 (3H,d), 1.99 (3H, s), 2.55 (1H, dd), 2.93 (1H, dd), 3.11-3.25 (1H, m), 4.77(2H, s), 6.35 (1H, dd), 6.52 (1H, dd), 6.81 (1H, t). Alkyl NH₂ protonsnot observed. m/z: ES+ [M+H]+ 165.

Preparation of(R)—N-(1-(3-amino-2-methylphenyl)propan-2-yl)-1-fluorocyclopropane-1-carboxamide

(R)-3-(2-aminopropyl)-2-methylaniline (1.70 g, 10.4 mmol) was dissolvedin DMF (29.9 mL) and treated with 1-fluorocyclopropane-1-carboxylic acid(1.00 g, 9.61 mmol), HATU (4.02 g, 10.6 mmol), and TEA (2.68 mL, 19.22mmol). The reaction was stirred at room temperature for 3 hours and thenquenched with water and extracted with EtOAc. The organic layer waswashed with saturated aqueous sodium chloride, dried over sodiumsulfate, and filtered. The filtrate was concentrated under reducedpressure, and the resulting residue was dried under vacuum overnight toremove residual DMF. The residue was then adsorbed onto silica andpurified by flash column silica chromatography, elution gradient 0 to80% ethyl acetate in hexanes to afford(R)—N-(1-(3-amino-2-methylphenyl)propan-2-yl)-1-fluorocyclopropane-1-carboxamide(1.47 g, 61.1%) as a light yellow solid. ¹H NMR (300 MHz, DMSO-d₆, 27°C.) 1.01-1.28 (7H, m), 2.02 (3H, s), 2.54-2.62 (1H, m), 2.83 (1H, dd),3.89-4.16 (1H, m), 4.68 (2H, s), 6.38 (1H, d), 6.49 (1H, d), 6.78 (1H,t), 8.14 (1H, d) m/z: ES+ [M+H]+251.

Preparation of(R)-3-(2-(((1-fluorocyclopropyl)methyl)amino)propyl)-2-methylaniline

Borane tetrahydrofuran complex in THF (1 M; 35.2 ml, 35.2 mmol) wasadded to a solution of(R)—N-(1-(3-amino-2-methylphenyl)propan-2-yl)-1-fluorocyclopropane-1-carboxamide(1.47 g, 5.87 mmol) in THF (13.7 mL) at room temperature under nitrogen.The reaction was then heated at 65° C. for 6 hrs. The reaction wascooled to 0° C. and quenched cautiously with MeOH (gas evolution). Thesolution was then concentrated under reduced pressure and stored in thefreezer for 18 hours. The residue was dissolved in MeOH (6 mL) andheated at 65° C. for 3 hrs. The solution was cooled to room temperatureand then concentrated under reduced pressure. The residue obtained waspurified by flash silica chromatography, elution gradient 0 to 16%methanol in DCM, to afford(R)-3-(2-(((1-fluorocyclopropyl)methyl)amino)propyl)-2-methylaniline(1.14 g, 82%) as colorless oil. ¹H NMR (300 MHz, DMSO-d₆, 27° C.)0.54-0.70 (2H, m), 0.79-1.02 (5H, m), 1.64 (1H, br. s.), 1.99 (3H, s),2.36 (1H, dd), 2.69-2.97 (4H, m), 4.68 (2H, s), 6.36 (1H, d), 6.49 (1H,d), 6.79 (1H, t). m/z: ES+ [M+H]+237.

Preparation of(1S,3R)-1-(5-bromopyridin-2-yl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine

5-Bromopicolinaldehyde (507 mg, 2.72 mmol) was added to a stirredsolution of(R)-3-(2-(((1-fluorocyclopropyl)methyl)amino)propyl)-2-methylaniline(322 mg, 1.36 mmol) in acetic acid (7028 μL) and water (143 μL). Theresulting mixture was heated to 90° C. and stirred at this temperaturefor 5 hours. The mixture was concentrated and dissolved in EtOAC (50 mL)and saturated NaHCO₃ (25 mL). The aqueous layer was extracted with EtOAc(2×50 mL). The combined organics were concentrated under reducedpressure. The residue was dissolved in DCM (20 mL) and 1M HCl (20 mL).The mixture was stirred vigorously for 30 minutes. The layers wereseparated and the the aqueous layer was washed with DCM (20 mL). Theaqueous layer was basified to >pH 10 with solid NaOH. The solution wasextracted with DCM (3×25 mL). The combined organics were dried (phaseseparator cartridge) and concentrated to give the crude product. Thecrude product was purified by flash silica chromatography, elutiongradient 0 to 60% EtOAc in heptane. Pure fractions were evaporated todryness to afford(1S,3R)-1-(5-bromopyridin-2-yl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(217 mg, 39%) as a yellow gum. ¹H NMR (500 MHz, CDCl₃, 27° C.) 0.33-0.47(1H, m), 0.49-0.6 (1H, m), 0.89-1.01 (2H, m), 1.04 (3H, d), 2.06 (3H,s), 2.55-2.64 (2H, m), 2.92-3.04 (2H, m), 3.51 (2H, s), 3.65-3.72 (1H,m), 4.88 (1H, s), 6.45 (1H, d), 6.58 (1H, d), 7.31 (1H, dd), 7.66 (1H,dd), 8.54 (1H, dd). m/z: ES+ [M+H]+ 404.

Preparation of(6S,8R)-6-(5-bromopyridin-2-yl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

(1S,3R)-1-(5-Bromopyridin-2-yl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(295 mg, 0.73 mmol) in propionic acid (8291 μL) was cooled to −20° C.Sodium nitrite (50.3 mg, 0.73 mmol) in water (829 μL) was added dropwiseover 2-3 minutes. The reaction mixture was stirred at −20° C. for 45minutes. The reaction mixture was diluted with ice-cold EtOAc (30 mL).The reaction mixture was poured into saturated NaHCO₃ (40 mL). Themixture was stirred vigorously for 5 minutes. The layers were separatedand the organic layer was washed with saturated NaHCO₃(2×30 mL). Thecombined aqueous layers were back extracted with EtOAc (2×30 mL). Thecombined organic phases were dried (Na₂SO₄) and concentrated to give thecrude product as brown oil. The crude material was purified by flashsilica column chromatography eluting with 0-45% EtOAc in heptane toafford(6S,8R)-6-(5-bromopyridin-2-yl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(107 mg, 35%) as an orange solid. ¹H NMR (500 MHz, CDCl₃, 27° C.)0.35-0.44 (1H, m), 0.51-0.63 (1H, m), 0.92-1.07 (2H, m), 1.09 (2H, d),2.69 (1H, dd), 2.96 (1H, dd), 3.01-3.10 (1H, m), 3.33-3.49 (2H, m),3.78-3.89 (1H, m), 5.03 (1H, s), 6.95 (1H, d), 7.19 (1H, d), 7.34-7.39(1H, m), 7.68 (1H, dd), 8.07 (1H, d), 8.58 (1H, dd). m/z: ES+ [M+H]+ 415and 417.

Preparation of(6S,8R)-6-(5-bromopyridin-2-yl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

3,4-Dihydro-2H-pyran (0.029 mL, 0.32 mmol) was added to a solution of(6S,8R)-6-(5-bromopyridin-2-yl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(107 mg, 0.26 mmol) and 4-methylbenzenesulfonic acid hydrate (9.80 mg,0.05 mmol) in DCM (2 mL) and the reaction was heated to 45° C. for 3hours. Further 3,4-dihydro-2H-pyran (0.235 mL, 2.58 mmol) and4-methylbenzenesulfonic acid hydrate (49.0 mg, 0.26 mmol) were added andthe mixture was heated at 45° C. for 16 hours. The reaction was dilutedwith DCM and washed with saturated NaHCO₃ solution. The organic phasewas dried and evaporated, then the crude was passed through a plug ofsilica (1:1 EtOAc/heptane). The filtrate was evaporated to afford(6S,8R)-6-(5-bromopyridin-2-yl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineas a brown oil that was used without further purification. m/z: ES+[M+H]+ 499.

Example 4N-(4-((6S,8R)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine(diastereoisomeric mixture)

1-(3-Fluoropropyl)azetidin-3-amine (50 mg, 0.37 mmol), a mixture of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineand(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-2H-pyrazolo[4,3-f]isoquinoline(140 mg, 0.25 mmol), cesium carbonate (163 mg, 0.50 mmol) and[(2-di-cyclohexylphosphino-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl)-2-(2′-amino-1,1′-biphenyl)]palladium(II)methanesulfonate (BrettPhos Pd G3) (23 mg, 0.02 mmol) were suspended in1,4-dioxane (2 mL) and sealed into a microwave tube. The reaction washeated to 100° C. for 4 hours under microwave irradiation. The reactionmixture was diluted with DCM (25 mL) and washed with water (25 mL). Theorganic layer was evaporated. The residue was dissolved in DCM (3 mL)and TFA (1 mL). The reaction mixture was stirred at room temperature for1 hour and then partitioned between DCM and 2M NaOH (20 mL each). Theorganic phase was evaporated and the crude product purified bypreparative HPLC (Waters CSH C18 OBD column, 5μ silica, 30 mm diameter,100 mm length), using decreasingly polar mixtures of water (containing1% NH₃) and MeCN as eluents to affordN-(4-((6S,8R)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine(26.0 mg, 20%) as a gum as a mixture of diastereoisomers. ¹H NMR (500MHz, DMSO-d₆, 27° C.) 0.94-0.99 (3H, m), 1.16-1.24 (3H, m), 1.64 (2H,dq), 2.39-2.47 (3H, m), 2.70 (2H, t), 2.74-2.84 (2H, m), 3.03-3.21 (2H,m), 3.24 (3H, d), 3.48 (2H, dd), 3.58-3.64 (2H, m), 3.79 (3H, d),3.88-3.94 (1H, m), 4.44 (2H, dt), 5.17 (1H, d), 5.88 (1H, td), 5.97 (1H,dd), 6.16 (1H, d), 6.32-6.38 (1H, m), 6.63 (1H, t), 7.16 (1H, dd), 8.02(1H, s), 12.91 (1H, s). ¹⁹F NMR (471 MHz, DMSO-d₆, 27° C.) −218.19,−149.21, −148.22. m/z: ES+[M+H]+ 528.

The mixture of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineand(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-2H-pyrazolo[4,3-f]isoquinolinewere prepared as follows;

Preparation of a diastereoisomeric mixture ofN—((R)-1-(3-bromo-2-methylphenyl)propan-2-yl)-2-fluoro-3-methoxy-2-methylpropan-1-amine

Trifluoromethanesulfonic anhydride (2.148 ml, 12.79 mmol) was added to((2-fluoro-3-methoxy-2-methylpropoxy)methanetriyl)tribenzene (4.44 g,12.18 mmol) dissolved in dichloromethane (50 mL) and the reactionmixture stirred at room temperature for 30 minutes. Triethylsilane(2.140 ml, 13.40 mmol) was then added and the mixture was stirred for afurther 30 minutes. The reaction was concentrated to afford cruderacemic 2-fluoro-3-methoxy-2-methylpropyl trifluoromethanesulfonate thatwas used directly in the next step without further purification.

DIPEA (5.36 mL, 30.68 mmol) was added to(R)-1-(3-bromo-2-methylphenyl)propan-2-amine (2.80 g, 12.3 mmol) and2-fluoro-3-methoxy-2-methylpropyl trifluoromethanesulfonate (3.12 g,12.3 mmol) in 1,4-dioxane (60 mL) at room temperature under nitrogen[exotherm]. The resulting mixture was stirred at 85° C. for 24 hours.The reaction mixture was allowed to cool and the reaction mixtureevaporated. The crude product was purified by flash silicachromatography, elution gradient 0 to 60% EtOAc in heptane to affordN—((R)-1-(3-bromo-2-methylphenyl)propan-2-yl)-2-fluoro-3-methoxy-2-methylpropan-1-amine(2.480 g, 73%) as an oil as a mixture of diastereoisomers. ¹H NMR (500MHz, CDCl₃, 27° C.) 1.02-1.07 (3H, m), 1.31 (1.5H, d), 1.32 (1.5H, d),2.41 (3H, s), 2.57-2.66 (1H, m), 2.68-2.90 (4H, m), 3.34 (1.5H, d), 3.35(1.5H, d), 3.38-3.46 (2H, m), 6.96 (1H, t), 7.07 (1H, d), 7.42 (1H, d).NH not seen. ¹⁹F NMR (471 MHz, CDCl₃, 27° C.) −157.94, −157.53. m/z: ES+[M+H]+332/334.

Preparation of a diastereoisomeric mixture of3-((R)-24(2-fluoro-3-methoxy-2-methylpropyl)amino)propyl)-2-methylaniline

Tris(dibenzylideneacetone)dipalladium(0) (0.205 g, 0.22 mmol) and(±)-2,2′-bis(diphenylphosphino)-1,1′-binaphthalene (0.279 g, 0.45 mmol)were added to a suspension of a diastereoisomeric mixture ofN—((R)-1-(3-bromo-2-methylphenyl)propan-2-yl)-2-fluoro-3-methoxy-2-methylpropan-1-amine(2.48 g, 7.46 mmol), benzophenone imine (1.487 g, 8.21 mmol) and sodiumtert-butoxide (1.076 g, 11.20 mmol) in degassed toluene (30 mL) and thereaction was heated to 90° C. for 3 hours. After cooling, the toluenewas evaporated. The residue was dissolved in DCM (250 mL) and washedwith water (250 mL). The aqueous was extracted with DCM (100 mL) and thecombined organics were concentrated to approximately 50 mL. 2 M HClsolution (50 mL) was added and the biphasic mixture was stirredvigorously for 30 minutes. The layers were separated and the aqueousphase was extracted with DCM. The aqueous phase was basified with 2 Maqueous NaOH. This was extracted with DCM (2×250 mL) and the combinedDCM extracts were evaporated to give3-((R)-2-((2-fluoro-3-methoxy-2-methylpropyl)amino)propyl)-2-methylaniline(2.00 g, 100%) as an oil as mixture of diastereoisomers. ¹H NMR (500MHz, CDCl₃, 27° C.) 1.04 (3H, d), 1.31 (1.5H, d), 1.32 (1.5H, d), 2.11(3H, s), 2.53-2.60 (1H, m), 2.70-2.89 (4H, m), 3.35 (2H, d), 3.36 (1.5H,d), 3.43 (1.5H, dd), 3.58 (2H, s), 6.57 (1H, d), 6.61 (1H, d), 6.94 (1H,t), 7.58 (1H, s). ¹⁹F NMR (471 MHz, CDCl₃, 27° C.) −157.61, −157.30.m/z: ES+ [M+H]+ 269.

Preparation of a diastereoisomeric mixture of(1S,3R)-1-(4-bromo-2-methoxyphenyl)-2-(2-fluoro-3-methoxy-2-methylpropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine

A diastereoisomeric mixture of3-((2R)-2-((2-Fluoro-3-methoxy-2-methylpropyl)amino)propyl)-2-methylaniline(2.00 g, 7.45 mmol) and 4-bromo-2-methoxybenzaldehyde (3.37 g, 15.6mmol) were heated in acetic acid (30 mL) and water (0.671 mL, 37.3 mmol)to 70° C. overnight. After cooling, the acetic acid was evaporated. Theresidue was dissolved in EtOAc (40 mL) and 2N HCl solution (40 mL) wasadded. The biphasic mixture was stirred for 30 minutes, then the layerswere separated. The organic phase was extracted with water, then thecombined aqueous phases were basified by addition of 2N NaOH solutionand extracted with DCM (2×200 mL). The combined DCM layers wereevaporated and the crude product was purified by flash silicachromatography, elution gradient 0 to 50% EtOAc in heptane to affordfirst(1S,3R)-1-(4-bromo-2-methoxyphenyl)-2-(2-fluoro-3-methoxy-2-methylpropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(0.354 g, 10%) as a single diastereoisomer. ¹H NMR (500 MHz, DMSO-d₆,27° C.) 0.91 (3H, d), 1.22 (3H, d), 1.94 (3H, s), 2.26-2.40 (2H, m),2.65-2.78 (2H, m), 3.16-3.25 (4H, m), 3.42 (1H, dd), 3.85 (3H, s), 4.61(2H, s), 5.10 (1H, s), 6.24 (1H, d), 6.37 (1H, d), 6.61 (1H, d), 6.95(1H, dd), 7.15 (1H, d). One proton partially obscured by water. ¹⁹F NMR(471 MHz, DMSO-d₆, 27° C.) −149.65. m/z: ES+ [M+H]+ 465/467. Followed byfractions containing a mixture of diastereoisomers of(1S,3R)-1-(4-bromo-2-methoxyphenyl)-2-(−2-fluoro-3-methoxy-2-methylpropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(1.700 g, 49%)¹⁹F NMR (471 MHz, DMSO-d₆, 27° C.) −149.65, −148.78. m/z:ES+ [M+H]+ 465/467. Followed by fractions containing the seconddiastereoisomer of(1S,3R)-1-(4-bromo-2-methoxyphenyl)-2-(2-fluoro-3-methoxy-2-methylpropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(0.274 g, 8%) as a single diastereoisomer. ¹H NMR (500 MHz, DMSO-d₆, 27°C.) 0.91 (3H, d), 1.16 (3H, d), 1.93 (3H, s), 2.26-2.4 (2H, m),2.63-2.74 (2H, m), 3.23 (3H, s), 3.35 (1H, dd), 3.47 (1H, dd), 3.85 (3H,s), 4.60 (2H, s), 5.09 (1H, s), 6.24 (1H, d), 6.37 (1H, d), 6.61 (1H,d), 6.93 (1H, dd), 7.15 (1H, d). One proton obscured by water. ¹⁹F NMR(471 MHz, DMSO-d₆, 27° C.) −148.78. m/z: ES+ [M+H]+ 465/467. All asgums.

Preparation of a diastereoisomeric mixture of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

A solution of sodium nitrite (0.277 g, 4.02 mmol) in water (1.0 mL) wasadded to a solution of a diastereoisomeric mixture of(1S,3R)-1-(4-bromo-2-methoxyphenyl)-2-(2-fluoro-3-methoxy-2-methylpropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(1.70 g, 3.65 mmol) in propionic acid (15 mL) at −10° C. The reactionwas stirred for 1 hour, then ice-cold EtOAc (20 mL) was added. Thereaction was quenched by addition of aqueous NaHCO₃ solution (30 mL) andstirred for 15 minutes, before being allowed to warm to roomtemperature. The organic phase was washed with aqueous NaHCO₃ solution(30 mL) and brine (20 mL), dried (Na₂SO₄), filtered and evaporated. Thecrude product was purified by flash silica chromatography, elutiongradient 0 to 50% EtOAc in heptane to afford(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.950 g, 55%) as a brown solid as a mixture of diastereoisomers. ¹H NMR(500 MHz, DMSO-d₆, 27° C.) 0.95-0.97 (3H, m), 1.14-1.23 (3H, m),2.31-2.40 (1H, m), 2.78-2.88 (2H, m), 3.11-3.21 (2.5H, s), 3.23 (1.5H,s), 3.32-3.38 (1H, m), 3.40-3.52 (2H, m), 3.88 (1.5H, s), 3.89 (1.5H,s), 5.28 (1H, s), 6.59-6.69 (2H, m), 6.92-6.96 (1H, m), 7.16-7.22 (2H,m), 8.05 (1H, s), 12.96 (1H, s). ¹⁹F NMR (471 MHz, DMSO-d₆, 27° C.)−149.65, −148.68. m/z: ES+[M+H]+ 476/478.

Preparation of a mixture of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineand(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-2H-pyrazolo[4,3-f]isoquinoline

4-Methylbenzenesulfonic acid hydrate (0.038 g, 0.20 mmol) was added to asolution of a diastereoisomeric mixture of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.950 g, 1.99 mmol) and 3,4-dihydro-2H-pyran (0.546 mL, 5.98 mmol) inDCM (30 mL) and the mixture was heated at 40° C. overnight. More3,4-dihydro-2H-pyran (0.546 mL, 5.98 mmol) and 4-methylbenzenesulfonicacid hydrate (0.038 g, 0.20 mmol) were added. Heating continued for 7hours. Further portions of 3,4-dihydro-2H-pyran (0.546 mL, 5.98 mmol)and 4-methylbenzenesulfonic acid hydrate (0.038 g, 0.20 mmol) were addedand the reaction heated overnight. The reaction mixture was diluted withDCM (50 mL) and washed with saturated aqueous NaHCO₃ (50 mL). Theorganic phase was evaporated to a dark brown oil and the crude productwas purified by flash silica chromatography, elution gradient 0 to 35%EtOAc in heptane to afford a mixture of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineand(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-2H-pyrazolo[4,3-f]isoquinoline(0.960 g, 86%) as a foam, as a mixture of diastereoisomers. ¹H NMR (471MHz, DMSO-d₆, 27° C.) −149.79, −149.74, −149.69, −149.65, −148.77,−148.73, −148.69, −148.66. m/z: ES+ [M+H]+ 560/562.

Example 53-((6S,8R)-6-(2,6-difluoro-4-((1-(3-fluoropropyl)azetidin-3-yl)amino)phenyl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)-2,2-difluoropropan-1-ol

TBAF in THF (1 M; 160 μL, 0.16 mmol) was added to a solution ofN-(4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)-1-(3-fluoropropyl)azetidin-3-amine(80 mg, 0.10 mmol) in THF (0.5 mL) at room temperature. After 4 hours,the reaction was concentrated under reduced pressure, and the resultingresidue was purified by flash silica chromatography, elution gradient 30to 90% (10% MeOH in DCM containing 1% NH₄OH) in DCM. Product fractionswere concentrated under reduced pressure, and the resulting residue wasrepurified using the same conditions. Product fractions were againconcentrated under reduced pressure, and the resulting residue waspurified by preparative HPLC (Waters XBridge Prep C18 OBD column, 5μsilica, 19 mm diameter, 100 mm length) using decreasingly polar mixturesof water (containing 0.2% ammonium hydroxide) and MeCN as eluents (40 to70% over 7 min) to afford3-((6S,8R)-6-(2,6-difluoro-4-((1-(3-fluoropropyl)azetidin-3-yl)amino)phenyl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)-2,2-difluoropropan-1-ol(17 mg, 31%) as white solid. ¹H NMR (500 MHz, DMSO-d₆, 27° C.) 1.02 (3H,d), 1.59-1.69 (2H, m), 2.44 (2H, t), 2.56-2.67 (1H, m), 2.72 (2H, t),2.88 (1H, dd), 3.02-3.13 (1H, m), 3.17 (1H, dd), 3.49-3.56 (1H, m),3.57-3.69 (3H, m), 3.91 (1H, tdt), 4.44 (2H, dt), 5.08 (1H, s), 5.25(1H, br. s), 6.07 (2H, d), 6.64 (1H, d), 6.69 (1H, d), 7.20 (1H, d),8.04 (1H, s), 12.95 (1H, br. s). One H not observed. m/z: ES+ [M+H]+524.

TheN-(4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)-1-(3-fluoropropyl)azetidin-3-aminewas prepared as follows:

Preparation 3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropan-1-ol

NaH in mineral oil (60 wt %; 343 mg, 8.58 mmol) was added in one portionto a stirred solution of 2,2-difluoropropane-1,3-diol (874 mg, 7.80mmol) in THF (32 mL) at 0° C. The reaction was allowed to warm to roomtemperature, and was stirred at room temperature for 2 h. The reactionmixture was again cooled to 0° C., and tert-butyldiphenylchlorosilane(2.0 mL, 7.8 mmol) was added dropwise via syringe. The reaction mixturewas allowed to warm to room temperature over 1 hour and was thenquenched with water and extracted with EtOAc. The organic layer wasdried with Na₂SO₄, filtered, and the filtrate was concentrated underreduced pressure. The resulting residue was purified by flash silicachromatography, eluting with isocratic 5% ethyl acetate in hexanes, toafford 3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropan-1-ol (1.94,71%) as a colorless oil. ¹H NMR (300 MHz, CHLOROFORM-d, 27° C.) δ ppm1.03-1.14 (9H, s), 3.87-3.93 (4H, m), 7.37-7.44 (6H, m), 7.64-7.66 (4H,m).

Preparation of 3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyltrifluoromethane sulfonate

A solution of 3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropan-1-ol(1.94 g, 5.55 mmol) and 2,6-dimethylpyridine (1.94 ml, 16.6 mmol) in DCM(18 ml) was cooled to −10° C. (salt/ice bath). Trifluoromethanesulfonicanhydride (1.88 ml, 11.1 mmol) was added slowly dropwise over 10minutes. The reaction was maintained under these conditions for 2 hours.The reaction was then washed with water, aqueous HCl (1N; 100 mL), andsaturated aqueous sodium bicarbonate. The organic layer was dried overMgSO₄, filtered, and concentrated under reduced pressure to afford3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyltrifluoromethanesulfonate (2.68 g, 100%) as a red oil. ¹H NMR (300 MHz,CHLOROFORM-d, 27° C.) 1.03-1.14 (9H, s), 3.90 (2H, t), 4.76 (2H, t),7.39-7.56 (6H, m), 7.59-7.75 (4H, m).

Preparation of 3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyltrifluoromethane sulfonate

3-((Tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyltrifluoromethanesulfonate (1.92 g, 3.98 mmol) was added to a solution of(R)-3-(2-aminopropyl)-2-methylaniline (0.784 g, 4.77 mmol) and DIPEA(1.031 ml, 5.97 mmol) in 1,4-dioxane (15 ml). The reaction was heated at85° C. for 18 hours. After cooling, the reaction was diluted with DCMand washed with water. The aqueous layer was extracted with DCM, and thecombined organic layers were dried over magnesium sulfate, filtered, andconcentrated under reduced pressure. The resulting residue was purifiedby flash silica chromatography, elution gradient 0 to 4% methanolicammonia in DCM. Pure fractions were concentrated to dryness to afford(R)-3-(2-((3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)amino)propyl)-2-methylaniline(1.97 g, 100%) as a yellow oil. ¹H NMR (300 MHz, METHANOL-d₄, 27° C.) δppm 0.97-1.12 (12H, m), 2.10 (3H, s), 2.53-2.63 (1H, m), 2.74-2.84 (1H,m), 2.86-2.99 (1H, m), 3.00-3.19 (2H, m), 3.80 (2H, td), 6.53 (1H, d),6.63 (1H, d), 6.86 (1H, t), 7.38-7.50 (6H, m), 7.64-7.72 (4H, m). m/z:ES+ [M+H]+ 497.

Preparation(1S,3R)-1-(4-bromo-2,6-difluorophenyl)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine

4-Bromo-2,6-difluorobenzaldehyde (1.55 g, 7.02 mmol) was added to asolution of(R)-3-(24(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)amino)propyl)-2-methylaniline(1.74 g, 3.51 mmol) in acetic acid (17 mL) and water (0.32 mL, 18 mmol),and the reaction was heated at 80° C. overnight. The reaction wasconcentrated under reduced pressure, and the residue was partitionedbetween EtOAc and saturated aqueous NaHCO₃. The organic layer wasconcentrated under reduced pressure and purified by flash silicachromatography, elution gradient 20 to 80% ethyl acetate in hexanes, toafford(1S,3R)-1-(4-bromo-2,6-difluorophenyl)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(1.33 g, 54.5%) as a solid.

¹H NMR (300 MHz, CHLOROFORM-d, 27° C.) δ ppm 1.04-1.11 (12H, m),2.25-2.38 (3H, m), 2.53-2.65 (1H, m), 2.73 (1H, q), 2.86-3.03 (1H, m),3.15-3.38 (1H, m), 3.52-3.71 (2H, m), 3.85-4.01 (1H, m), 5.31 (1H, d),6.58-6.64 (1H, m), 6.67-6.73 (1H, m), 6.88-6.95 (2H, m), 7.18-7.24 (2H,m), 7.37-7.50 (6H, m), 7.60-7.70 (4H, m). m/z: ES+ [M+H]+ 699.

Preparation of(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

Sodium nitrite (0.130 g, 1.89 mmol) in water (0.900 mL) was addeddropwise to a solution of(1S,3R)-1-(4-bromo-2,6-difluorophenyl)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(1.32 g, 1.89 mmol) in propionic acid (9 mL) at −8° C. (salt/ice bath)and stirred under these conditions for 20 minutes. The reaction wasdiluted with ethyl acetate (20 mL, precooled to −10° C.) and quenchedwith slow addition of saturated aqueous NaHCO₃(30 mL, precooled to −10°C.) over 15 min at 0° C. The mixture was allowed to warm to roomtemperature and maintained under these conditions for 18 hours. Theorganic layer was separated, and the aqueous layer was extracted withEtOAc. The combined organic layers were washed with saturated aqueousNaHCO₃ and water, dried over sodium sulfate, filtered, and concentratedunder reduced pressure. The resulting residue was purified by flashsilica chromatography, elution gradient 15 to 50% ethyl acetate inhexanes, to afford(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.520 g, 38.8%) as light brown solid. ¹H NMR (300 MHz, CHLOROFORM-d,27° C.) δ ppm 1.07 (9H, s), 1.12 (3H, d), 2.72-2.92 (2H, m), 3.21-3.39(2H, m), 3.51-3.66 (1H, m), 3.67-3.79 (1H, m), 3.86-4.00 (1H, m), 5.37(1H, s), 6.81 (1H, d), 6.87-6.98 (2H, m), 7.25-7.29 (1H, m), 7.35-7.50(6H, m), 7.59-7.70 (4H, m), 8.14 (1H, s). Indole NH not observed. m/z:ES+ [M+H]+ 710.

Preparation of(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

3,4-Dihydro-2H-pyran (95 μL, 1.04 mmol) was added to a solution of(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(492 mg, 0.69 mmol) and para-toluenesulfonic acid monohydrate (13 mg,0.07 mmol) in DCM (3.5 mL), and the reaction was heated under refluxconditions for 6 hours. After cooling, the reaction was washed withsaturated aqueous NaHCO₃, dried over Na₂SO₄, filtered, and concentratedunder reduced pressure. The residue was purified by flash silicachromatography, elution gradient 10 to 30% ethyl acetate in hexanes, toafford(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(474 mg, 86%) as a light brown gummy solid. ¹H NMR (300 MHz,CHLOROFORM-d, 27° C.) 1.06 (9H, s), 1.19-1.25 (3H, m), 1.74-1.84 (3H,m), 2.09-2.20 (2H, m), 2.49-2.67 (1H, m), 2.84-2.96 (2H, m), 3.27-3.45(2H, m), 3.61-3.87 (3H, m), 4.04 (2H, d), 5.52 (1H, s), 5.66-5.72 (1H,m), 6.78 (1H, d), 6.94 (2H, d), 7.32 (1H, d), 7.36-7.50 (6H, m),7.60-7.68 (4H, m), 8.04 (1H, s).

Preparation of tert-butyl3-((4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)amino)azetidine-1-carboxylate

A degassed mixture of(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.474 g, 0.600 mmol), tert-butyl 3-aminoazetidine-1-carboxylate (0.15g, 0.89 mmol), Xantphos (0.069 g, 0.12 mmol), Pd₂(dba)₃ (0.06 g, 0.06mmol), and cesium carbonate (0.389 g, 1.19 mmol) in dioxane (3 mL) washeated at 115° C. for 4 hours. The reaction was then concentrated underreduced pressure, and the resulting residue was adsorbed onto silica geland purified by flash silica chromatography, elution gradient 5 to 50%ethyl acetate in hexanes, to afford tert-butyl3-((4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)amino)azetidine-1-carboxylate(0.377 g, 71.3%) as a solid. ¹H NMR (300 MHz, CHLOROFORM-d, 27° C.) 1.06(9H, s), 1.12 (3H, d), 1.46 (9H, s), 1.65-1.84 (3H, m), 2.07-2.23 (2H,m), 2.58 (1H, d), 2.75-2.91 (2H, m), 3.16-3.34 (2H, m), 3.52-3.78 (5H,m), 3.91-4.07 (3H, m), 4.19-4.29 (2H, m), 5.23-5.34 (1H, m), 5.67 (1H,dd), 5.81 (2H, d), 6.81 (1H, d), 7.25-7.29 (1H, m), 7.35-7.48 (6H, m),7.65 (4H, ddd), 8.02 (1H, s). Aniline NH not observed. m/z: ES+ [M+H]+886.

Preparation ofN-(4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)azetidin-3-amine

TFA (328 μL, 4.25 mmol) was added slowly to a solution of tert-butyl3-((4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin6 yl) 3,5-difluorophenyl)amino)azetidine-1-carboxylate (377 mg, 0.43mmol) in DCM (2 mL) at 0° C. The reaction was allowed to warm to roomtemperature and was stirred under these conditions for 1 hour. Thereaction was then concentrated under reduced pressure, and the resultingresidue was partitioned between ethyl acetate and saturated aqueousNaHCO₃. The organic layer was dried over MgSO₄, filtered, andconcentrated under reduced pressure. The resulting residue was purifiedby flash silica chromatography, elution gradient 30 to 70% (10% methanolin DCM containing 1% ammonium hydroxide) in DCM to affordN-(4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)azetidin-3-amine(166 mg, 55.6%) as pale solid. ¹H NMR (300 MHz, CHLOROFORM-d, 27° C.)0.96-1.20 (12H, m), 2.62-3.03 (2H, m), 3.10-3.42 (2H, m), 3.42-3.63 (3H,m), 3.63-3.80 (2H, m), 3.81-4.08 (3H, m), 4.15-4.28 (1H, m), 4.38 (1H,d), 5.23 (1H, s), 5.83 (2H, m), 6.80 (1H, m), 7.15 (1H, d), 7.32-7.53(6H, m), 7.61-7.81 (4H, m), 8.06 (1H, s). Indazole NH not observed. m/z:ES+ [M+H]+ 702.

Preparation ofN-(4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)-1-(3-fluoropropyl)azetidin-3-amine

1-Fluoro-3-iodopropane (40 mg, 0.21 mmol) was added to a solution ofN-(4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)azetidin-3-amine(144 mg, 0.21 mmol) and DIPEA (107 μL, 0.62 mmol) in DMF (1 mL) atambient temperature. The reaction was stirred at room temperature fo 3hours and then diluted with EtOAc (40 mL) and washed with saturatedaqueous sodium chloride (3×20 mL). The organic layers were dried overMgSO₄, filtered, and concentrated under reduced pressure. The resultingresidue was purified by flash silica chromatography, elution gradient 15to 30% (10% methanol in DCM containing 1% ammonium hydroxide) in DCM, toaffordN-(4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)-1-(3-fluoropropyl)azetidin-3-amine(87 mg, 56%) as yellow solid. m/z: ES+ [M+H]+ 762.

Example 6N-(4-((6S,8R)-7-((l-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine

1-Fluoro-3-iodopropane (40.9 μL, 0.39 mmol) was added to a solution ofN-(4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)azetidin-3-amine(169 mg, 0.39 mmol) and DIPEA (203 μL, 1.16 mmol) in NMP (1.7 mL) atroom temperature. After 18 hours, the reaction was concentrated underreduced pressure. The resulting residue was purified by reverse phaseflash chromatography (C18), eluting with decreasingly polar mixtures ofwater (containing 0.2% ammonium hydroxide) and MeCN as eluents. Productfractions were combined and lyopholized to affordN-(4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine(75 mg, 39%) as clear residue. ¹H NMR (500 MHz, DMSO-d₆, 27° C.)0.39-0.46 (1H, m), 0.47-0.54 (1H, m), 0.77-0.85 (2H, m), 0.91 (3H, d),1.51-1.64 (2H, m), 2.38 (2H, t), 2.54 (1H, dd), 2.63 (2H, q), 2.75-2.90(2H, m), 3.14 (1H, dd), 3.52-3.64 (3H, m), 3.73 (3H, s), 3.79-3.88 (1H,m), 4.38 (2H, dt), 5.10 (1H, s), 5.82-5.87 (1H, m), 5.91 (1H, d), 6.09(1H, d), 6.49 (1H, d), 6.57 (1H, d), 7.07 (1H, d), 7.94 (1H, s), 12.83(1H, s). m/z: ES+ [M+H]+ 496.

TheN-(4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)azetidin-3-aminewas prepared as follows.

Preparation of(1S,3R)-1-(4-bromo-2-methoxyphenyl)-24(1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine

(R)-3-(2-(((1-Fluorocyclopropyl)methyl)amino)propyl)-2-methylaniline(451 mg, 1.91 mmol) and 4-bromo-2-methoxybenzaldehyde (410 mg, 1.91mmol) in a mixture of water (172 mg, 9.54 mmol) and acetic acid (7.5 mL)were heated at 80° C. for 6 hours. The reaction was concentrated underreduced pressure, and the resulting residue was treated with aqueous HCl(1N; 18 mL). The mixture was stirred at 80° C. for 18 hours. Thereaction was then cooled and saturated aqueous NaHCO₃ was added. Themixture was extracted with DCM, and the organic layer was concentratedunder reduced pressure. The resulting residue was purified flash silicachromatography, elution gradient 5 to 35% ethyl acetate in hexanes, toafford(1S,3R)-1-(4-bromo-2-methoxyphenyl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(591 mg, 71.5%) as a gummy film. ¹H NMR (300 MHz, DMSO-d₆, 27° C.)0.42-0.61 (2H, m), 0.81-0.91 (2H, m), 0.93 (3H, d), 1.93 (3H, s),2.43-2.48 (2H, m), 2.72-2.89 (2H, m), 3.48-3.57 (1H, m), 3.87 (3H, s),4.57 (2H, s), 5.12 (1H, s), 6.24 (1H, d), 6.35 (1H, d), 6.82 (1H, d),6.96 (1H, d), 7.16 (1H, d). m/z: ES+ [M+H]+ 433.

Preparation of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

Acetic acid (0.392 ml, 6.84 mmol) was added to(1S,3R)-1-(4-bromo-2-methoxyphenyl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(0.593 g, 1.37 mmol) in CHCl₃ (5 mL), and the resulting solution wascooled to 0° C. Isopentyl nitrite (0.321 g, 2.74 mmol) in CHCl₃ (1 mL)was added to the reaction dropwise, and the reaction was stirred at 0°C. for another 2 hours. Then saturated aqueous NaHCO₃ (1 g in 20 mL ofwater) was added slowly. The reaction was stirred at 0° C. for 10minutes, and then the layers were separated. The organic layer waspurified by flash column chromatography, elutiong gradient 5 to 35%ethyl acetate in hexanes, to afford(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.299 g, 49.2%) as light brown solid. ¹H NMR (300 MHz, DMSO-d₆, 27° C.)δ ppm 0.39-0.65 (2H, m), 0.77-0.96 (2H, m), 1.00 (3H, d), 2.52-2.63 (1H,m), 2.87-3.03 (2H, m), 3.21-3.28 (1H, m), 3.71 (1H, d), 3.92 (3H, s),5.31 (1H, s), 6.65 (1H, d), 6.86-6.92 (1H, m), 6.97 (1H, m), 7.18 (1H,d), 7.23 (1H, d), 8.05 (1H, s), 12.94 (1H, s). m/z: ES+ [M+H]+ 444.

Preparation of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

3,4-Dihydro-2H-pyran (176 μL, 1.93 mmol) was added to a solution of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(286 mg, 0.64 mmol) and para-toluenesulfonic acid monohydrate (12 mg,0.060 mmol) in DCM (2.5 ml). The reaction was subjected to microwaveconditions (100° C., 300 W) for 6 hours. Upon cooling, the reaction waswashed with saturated aqueous NaHCO₃, dried over Na₂SO₄, filtered, andconcentrated under reduced pressure. The resulting residue was purifiedby flash silica chromatography, elution gradient 10 to 30% ethyl acetatein hexanes, to afford(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(299 mg, 88%) as a light brown gummy solid. m/z: ES+ [M+H]+528.

Preparation of tert-butyl3-((4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)amino)azetidine-1-carboxylate

(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(278 mg, 0.53 mmol), tert-butyl 3-aminoazetidine-1-carboxylate (136 mg,0.79 mmol), cesium carbonate (343 mg, 1.05 mmol), and BrettPhos 3rdGeneration Precatalyst (24 mg, 0.030 mmol) in dioxane (2.5 mL) weresubjected to microwave conditions (100° C., 300 W) for 6 hours. Thereaction was then concentrated under reduced pressure, and the resultingresidue was adsorbed onto silica gel before purification via flashsilica chromatography, elution gradient 20 to 60% ethyl acetate inhexanes, to afford tert-butyl3-((4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)amino)azetidine-1-carboxylate(296 mg, 91%). m/z: ES+ [M+H]+ 620.

Preparation ofN-(4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)azetidin-3-amine

HCl in dioxane (4 M; 1.2 mL, 4.78 mmol) was added dropwise to a solutionof tert-butyl3-((4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)amino)azetidine-1-carboxylate(296 mg, 0.48 mmol) in MeOH (3.5 ml). After stirring at room temperatureovernight, the reaction was concentrated under reduced pressure, and theresulting residue was dissolved in MeOH (2 mL). This solution was thenloaded onto an SCX-2 ion-exchange cartridge that had been pretreatedwith methanol. The cartridge was washed with methanol and then 7Nammonia in methanol. Product fractions were concentrated under reducedpressure to affordN-(4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)azetidin-3-amine(181 mg, 87%) as a pale solid. m/z: ES+ [M+H]+ 436.

Example 7(6S,8R)-7-((1-fluorocyclopropyl)methyl)-6-(4-((1-(3-fluoropropyl)azetidin-3-yl)oxy)-2-methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

1-Fluoro-3-iodopropane (5.79 μL, 0.05 mmol) was added to a solution of(6S,8R)-6-(4-(azetidin-3-yloxy)-2-methoxyphenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolinetrifluoroacetic acid salt (0.030 g, 0.050 mmol) and DIPEA (0.029 mL,0.16 mmol) in NMP (0.52 ml) at room temperature. After 3 hours, thereaction was diluted with EtOAc (40 mL) and washed with saturatedaqueous sodium chloride (3×20 mL). The organic layer was dried overMgSO₄, filtered and concentrated under reduced pressure. The resultingresidue was purified by flash silica chromatography, elution gradient 15to 30% (MeOH in DCM containing 1% ammonium hydroxide) in DCM to afford(6S,8R)-7-((1-fluorocyclopropyl)methyl)-6-(4-((1-(3-fluoropropyl)azetidin-3-yl)oxy)-2-methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(5 mg, 18%) as white solid. ¹H NMR (500 MHz, Methanol-d₄, 26° C.)0.45-0.56 (2H, m), 0.85-1.03 (2H, m), 1.12 (3H, d), 1.67-1.81 (2H, m),2.50-2.62 (1H, m), 2.61-2.68 (2H, m), 2.92 (1H, dd), 3.08-3.21 (3H, m),3.25-3.28 (1H, m), 3.70-3.84 (3H, m), 3.86-3.91 (3H, m), 4.45 (2H, dt),4.73-4.82 (1H, m), 5.49 (1H, br. s), 6.19 (1H, d), 6.50 (1H, br. s.),6.72 (1H, d), 6.79 (1H, d), 7.18 (1H, d), 8.05 (1H, s). Indazole NH notobserved. m/z: ES+[M+H]+ 497.

TheN-(4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)azetidin-3-aminetrifluoroacetic acid salt was prepared as follows:

Preparation of(6S,8R)-7-((l-fluorocyclopropyl)methyl)-6-(2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

A mixture of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(369 mg, 0.70 mmol; prepared according to Example 6),bis(pinacolato)diboron (266 mg, 1.05 mmol),[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (51 mg,0.070 mmol) and potassium acetate (233 mg, 2.37 mmol) in dioxane (3.50mL) was degassed and purged with nitrogen. The reaction mixture was thenwarmed to 80° C. After stirring under these conditions for 18 hours, thereaction mixture was cooled to room temperature, filtered through a padof Celite®, and washed with EtOAc(100 mL). The filtrate was washed withwater (100 mL), the aqueous layer was extracted with EtOAc (2×70 mL),and the combined organic layers were washed with water (3×70 mL) andsaturated aqueous sodium chloride (50 mL) before being dried over MgSO₄,filtered, and concentrated under reduced pressure. The resulting residuewas purified by flash silica chromatography, elution gradient 0 to 25%ethyl acetate in hexane to(6S,8R)-7-((1-fluorocyclopropyl)methyl)-6-(2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(186 mg, 46.3%) as a yellow solid. m/z: ES+ [M+H]+576.

Preparation of4-((6S,8R)-7-((l-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenol

Aqueous hydrogen peroxide (33%; 75 μL, 0.74 mmol) was added dropwise toa stirred solution of(6S,8R)-7-((1-fluorocyclopropyl)methyl)-6-(2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(186 mg, 0.32 mmol) and aqueous sodium hydroxide (1 M; 350 μL, 0.32mmol) in THF (2.5 mL) at 5° C. under air. The resulting mixture wasstirred at 5° C. for 5 minutes. The reaction was then diluted with water(50 mL) and DCM (100 mL). The layers were separated, and the aqueouslayer was extracted with DCM (2×50 mL). The combined organic layers weredried over sodium sulfate, filtered, and concentrated under reducedpressure. The resulting residue was purified by flash silicachromatography, elution gradient 20 to 50% ethyl acetate in hexanes, toafford4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-j]isoquinolin-6-yl)-3-methoxyphenol(94 mg, 62.5%) as a yellow solid. m/z: ES+ [M+H]+ 466.

Preparation of tert-butyl3-(4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenoxy)azetidine-1-carboxylate

Diethyl (E)-diazene-1,2-dicarboxylate (88 mg, 0.20 mmol) was added to asolution of4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenol(94 mg, 0.20 mmol), tert-butyl 3-hydroxyazetidine-1-carboxylate (35 mg,0.20 mmol), triphenylphosphine (53 mg, 0.20 mmol) and toluene (2.03 mL).The reaction was then warmed to 110° C. After 15 hours, the reaction wascooled, concentrated under reduced pressure, and purified by flashsilica chromatography, elution gradient 0 to 30% ethyl acetate inhexanes, to afford tert-butyl3-(4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenoxy)azetidine-1-carboxylate(95 mg, 75%). m/z: ES+ [M+H]+ 621.

Preparation of tert-butyl3-(4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenoxy)azetidine-1-carboxylatetrifluoroacetic acid salt

HCl in dioxane (4N; 383 μL, 1.53 mmol) was added dropwise to a solutionof tert-butyl3-(4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenoxy)azetidine-1-carboxylate(95 mg, 0.15 mmol) in MeOH (1 mL), and the reaction was stirred at roomtemperature for 18 hours. The reaction was concentrated under reducedpressure, and the resulting residue was purified by reverse phase flashchromatography, using decreasingly polar mixtures of water (containing0.01% TFA) and MeCN as eluents (20 to 80%) to afford(6S,8R)-6-(4-(azetidin-3-yloxy)-2-methoxyphenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(30 mg, 36%) as a TFA salt. ¹H NMR (300 MHz, METHANOL-d₄, 27° C.)0.86-1.09 (2H, m), 1.26-1.52 (3H, m), 1.59 (3H, d), 3.22-3.31 (1H, m),3.36-3.47 (1H, m), 3.66 (1H, dd), 4.03 (3H, s), 4.07-4.27 (4H, m),4.52-4.62 (2H, m), 5.14-5.27 (1H, m), 6.33 (1H, dd), 6.57-6.63 (1H, m),6.68 (1H, d), 6.75 (1H, s), 6.93 (1H, d), 7.50 (1H, d), 8.23 (1H, s).Indazole NH and addition H not observed. m/z: ES+ [M+H]+ 437.

Example 8N-(3,5-difluoro-4-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine

1-Fluoro-3-iodopropane (5.4 μl, 0.050 mmol) and diisopropylethylamine(0.013 mL, 0.070 mmol) were added to a stirred solution ofN-(3,5-difluoro-4-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-amine(22 mg, 0.050 mmol) in NMP (0.4 mL) at ambient temperature. After 16hours the crude reaction was purified directly by reverse phasepreparative HPLC (Waters XBridge C18 column, 19 mm diameter, 100 mmlength, 5 μm silica), elution gradient 40 to 70% MeCN in watercontaining 0.2% ammonium hydroxide as a modifier. Product fractions wereconcentrated to dryness to affordN-(3,5-difluoro-4-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine(9 mg, 36%) as a pale yellow solid. ¹H NMR (500 MHz, CD₂Cl₂, 27° C.)1.11 (3H, d), 1.69-1.78 (3H, m), 2.51-2.59 (2H, m), 2.81-2.94 (3H, m),2.95-3.03 (1H, m), 3.16-3.30 (1H, m), 3.59-3.70 (3H, m), 3.94-4.05 (1H,m), 4.41 (1H, t), 4.45 (1H, d), 4.51 (1H, t), 5.27 (1H, s), 6.02 (2H,d), 6.83 (1H, d), 7.21 (1H, d), 8.03 (1H, s), 10.47 (1H, br. s). m/z:ES+ [M+H]+ 512.

TheN-(3,5-difluoro-4-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-aminewas prepared as follows;

Preparation of of 2,2,2-trifluoroethyl trifluoromethanesulfonate

Trifluoromethanesulfonic anhydride (3.14 mL, 18.6 mmol) was addeddropwise via syringe over 5 minutes to a stirred solution of2,2,2-trifluoroethan-1-ol (1.23 mL, 16.9 mmol) and 2,6-dimethylpyridine(2.36 mL, 20.3 mmol) in DCM (50 mL) at −10° C. After 2 hours thereaction was washed successively with aqueous HCl (1N; 2×30 mL) andsaturated aqueous NaHCO₃ (20 mL). The organic layer was then dried overMgSO₄, filtered, and concentrated under reduced pressure to give2,2,2-trifluoroethyl trifluoromethanesulfonate (0.92 g, 23%) as a redoil. ¹H NMR (300 MHz, CDCl₃, 27° C.) 4.69 (2H, q).

Preparation of(R)-2-methyl-3-(2-((2,2,2-trifluoroethyl)amino)propyl)aniline

2,2,2-Trifluoroethyl trifluoromethanesulfonate (1.3 g, 2.8 mmol) wasadded to a stirred solution of (R)-3-(2-aminopropyl)-2-methylaniline(0.460 g, 2.80 mmol; prepared according to Example 3) anddiisopropylethylamine (0.636 mL, 3.64 mmol) in 1,4-dioxane (10 mL). Thereaction was heated at 65° C. for 15 hours and then cooled andconcentrated under reduced pressure. The resulting residue was dissolvedin EtOAc (50 mL) and washed with a mixture of saturated aqueous NaHCO₃and saturated aqueous sodium chloride. The aqueous layer was extractedwith EtOAc (20 mL), and the combined organic layers were dried overMgSO₄, filtered, and concentrated under reduced pressure. The resultingresidue was dissolved in MeOH, adsorbed onto diatomaceous earth underreduced pressure, and purified by flash silica chromatography, elutiongradient 1 to 6% MeOH in DCM. Product fractions were concentrated todryness to afford(R)-2-methyl-3-(2-((2,2,2-trifluoroethyl)amino)propyl)aniline (0.30 g,44%) as a pale orange gum. ¹H NMR (300 MHz, CDCl₃, 27° C.) 1.07 (3H, d),2.09 (3H, s), 2.63 (1H, dd), 2.76 (1H, dd), 2.92-3.05 (1H, m), 3.15 (2H,q), 6.58 (2H, d), 6.85-7.00 (1H, m). Signals for three NH's notobserved. m/z: ES+ [M+H]+ 247.

Preparation of(1S,3R)-1-(4-bromo-2,6-difluorophenyl)-3,5-dimethyl-2-(2,2,2-trifluoroethyl)-1,2,3,4-tetrahydroisoquinolin-6-amine

(R)-2-methyl-3-(2-((2,2,2-trifluoroethyl)amino)propyl)aniline (303 mg,1.23 mmol) and 4-bromo-2,6-difluorobenzaldehyde (544 mg, 2.46 mmol) wereheated in a mixture of acetic acid (5 mL) and water (0.111 mL) at 90° C.for 2 hours. The reaction was allowed to slowly cool to ambienttemperature and stirred overnight. The reaction was then concentratedunder reduced pressure, and the resulting residue was dissolved in EtOAc(30 mL) and washed with saturated aqueous NaHCO₃ (2×20 mL). The combinedaqueous layers were extracted with EtOAc (2×20 mL), and the combinedorganic layers were washed with saturated aqueous sodium chloride andconcentrated to minimal volume (˜20 mL). Aqueous HCl (1N; 20 mL) wasadded, and the biphasic mixture was stirred vigorously for 2 hours. Thelayers were separated, and the aqueous layer was extracted with EtOAc(20 mL). The aqueous layer was basified by the addition of solid sodiumcarbonate (until pH ˜8 as measured using a pH strip) and then extractedwith DCM (3×20 mL). The combined DCM extracts were dried over MgSO₄,filtered, and concentrated under reduced pressure to afford 40 mg ofcrude product. Meanwhile, the combined EtOAc extracts were washed withsaturated aqueous sodium chloride and concentrated to dryness. Theresultant residue was dissolved in THF (10 mL) and stirred withpolystyrene-bound tosylhydrazide (1.03 g, 2.76 mmol) for 14 hours. Themixture was filtered, and the resin was washed successively with THF (10mL) and MeOH (3×10 mL). The filtrate was combined with the previouslyobtained ca. 40 mg of crude product and concentrated under reducedpressure. This final residue was purified by flash silicachromatography, elution gradient 0 to 30% EtOAc in hexanes. Productfractions were concentrated to dryness to afford(1S,3R)-1-(4-bromo-2,6-difluorophenyl)-3,5-dimethyl-2-(2,2,2-trifluoroethyl)-1,2,3,4-tetrahydroisoquinolin-6-amine(230 mg, 41%) as a pale yellow solid. ¹H NMR (300 MHz, CDCl₃, 27° C.)1.04 (3H, d), 2.42 (3H, s), 2.58 (1H, dd), 2.74-2.87 (1H, m), 3.02-3.28(2H, m), 3.46-3.59 (1H, m), 5.27 (1H, s), 6.66 (1H, d), 7.02 (2H, d),7.32 (1H, d), 9.95-10.73 (2H, br. s). m/z: ES+[M+H]+ 449.

Preparation of(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

A solution of sodium nitrite (35 mg, 0.51 mmol) in water (0.20 mL) wasadded dropwise over 1 minute to a stirred solution of(1S,3R)-1-(4-bromo-2,6-difluorophenyl)-3,5-dimethyl-2-(2,2,2-trifluoroethyl)-1,2,3,4-tetrahydroisoquinolin-6-amine(230 mg, 0.51 mmol) in propionic acid (2 mL) at approximately −20° C.(ice-NaCl bath). After 15 minutes the reaction was diluted with ice-coldEtOAc (15 mL). The biphasic mixture was stirred vigorously under theseconditions and neutralized by the slow addition of solid Na₂CO₃ (untilbasic as measured using a pH strip). The cooling bath was removed. Thephases were separated, and the organic layer washed with saturatedaqueous NaHCO₃ (2×15 mL), saturated aqueous sodium chloride (15 mL),dried over MgSO₄, filtered, and concentrated under reduced pressure. Theresulting residue was purified by flash silica chromatography, elutiongradient 5 to 35% EtOAc in hexanes. Product fractions were concentratedto dryness to afford(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(104 mg, 45%) as a pale orange film. ¹H NMR (300 MHz, CDCl₃, 27° C.)1.15 (3H, d), 2.89-3.04 (2H, m), 3.20-3.36 (1H, m), 3.50 (1H, dd),3.61-3.75 (1H, m), 5.41 (1H, s), 6.80 (1H, d), 7.01-7.11 (2H, m), 7.23(1H, d), 8.10 (1H, s), 10.24 (1H, br. s). m/z: ES+ [M+H]+ 460.

Preparation of(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

3,4-Dihydro-2H-pyran (0.103 mL, 1.13 mmol) and p-toluenesulfonic acidmonohydrate (2 mg, 0.01 mmol) were added to a stirred solution of(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(104 mg, 0.23 mmol) in DCM (1.5 mL). The reaction was maintained underreflux conditions for 1 hour. After cooling, the reaction was dilutedwith DCM, washed with saturated aqueous NaHCO₃, dried over MgSO₄,filtered, and concentrated under reduced pressure. The resulting residuewas purified by flash silica chromatography, elution gradient 0 to 30%EtOAc in hexanes. Product fractions were concentrated to dryness toafford(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(117 mg, 95%) as pale yellow solid. m/z: ES+ [M+H]+ 544.

Preparation of tert-butyl3-((3,5-difluoro-44(6S,8R)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)amino)azetidine-1-carboxylate

A vial was charged with a stir bar,(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(55 mg, 0.10 mmol), tert-butyl 3-aminoazetidine-1-carboxylate (26 mg,0.15 mmol), Pd₂dba₃ (9 mg, 0.01 mmol), Xantphos (12 mg, 0.02 mmol), andcesium carbonate (99 mg, 0.30 mmol). The vial was sealed andevacuated/backfilled with nitrogen (3×) prior to the addition ofdegassed 1,4-dioxane (1 mL) via syringe. The mixture was stirred atambient temperature for 2 minutes then placed in a heating block at thathad been pre-heated to 90° C. After 22 hours, the mixture was allowed tocool to room temperature and then diluted with EtOAc. The mixture wasfiltered through diatomaceous earth, and the filtrate was concentratedunder reduced pressure. The resulting residue was purified by flashsilica chromatography, elution gradient 0 to 45% EtOAc in hexanes.Product fractions were concentrated to dryness to afford tert-butyl3-((3,5-difluoro-4-((6S,8R)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)amino)azetidine-1-carboxylate(46 mg, 72%) as a pale yellow solid. ¹H NMR (300 MHz, CDCl₃, 27° C.)1.08 (3H, d), 1.42 (9H, s), 1.55-1.81 (4H, m), 1.97-2.23 (2H, m),2.45-2.63 (1H, m), 2.80-3.02 (2H, m), 3.09-3.27 (1H, m), 3.34-3.47 (1H,m), 3.53-3.65 (1H, m), 3.66-3.75 (3H, m), 3.92-4.02 (1H, m), 4.19-4.32(3H, m), 5.25 (1H, s), 5.58-5.67 (1H, m), 5.93 (2H, dd), 6.81 (1H, d),7.22-7.27 (1H, m), 7.98 (1H, s). Partial overlap of the multiplet at7.22 to 7.27 ppm with chloroform. m/z: ES+ [M+H]+ 636.

Preparation ofN-(3,5-difluoro-4-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-amine

The tert-butyl3-((3,5-difluoro-4-((6S,8R)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)amino)azetidine-1-carboxylate(46 mg, 0.08 mmol) was dissolved in formic acid (0.50 mL, 13 mmol), andthe stirred solution warmed to 30° C. After 27 hours the reaction wasconcentrated under reduced pressure. The resulting residue was dissolvedin 5% IPA/DCM and neutralized with saturated aqueous NaHCO₃. The phaseswere separated, and the aqueous layer was extracted with 5% IPA/DCM (2×4mL). The combined organic layers were dried over MgSO₄, filtered, andconcentrated under reduced pressure to afford crudeN-(3,5-difluoro-4-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-amine(38 mg, 111%) contaminated with a small amount of partially deprotectedstarting material as a pale yellow film. m/z: ES+[M+H]+ 452.

Example 9

5-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)-4-methoxypyridin-2-amine

DMF (1 mL) and DIPEA (0.022 ml, 0.13 mmol) were added sequentially to aflask charged withN-(azetidin-3-yl)-5-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-j]isoquinolin-6-yl)-4-methoxypyridin-2-amine(22 mg, 0.050 mmol). 1-Fluoro-3-iodopropane (9 mg, 0.05 mmol) in DMF(0.1 mL) was then added. After 2 hours, the reaction was diluted withsaturated aqueous sodium chloride, and the mixture was extracted inEtOAC (3×). The combined organic layers were washed with water and driedover sodium sulfate, filtered and concentrated under reduced pressure.The resulting crude film was purified by flash silica chromatography,elution gradient 2 to 10% MeOH in DCM, to give5-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)-4-methoxypyridin-2-amine(9.0 mg, 36%) as a dry film. ¹H NMR (500 MHz, CHLOROFORM-d, 27° C.) δppm 0.46-0.66 (2H, m), 0.85-1.05 (2H, m), 1.09 (3H, d), 1.64-1.78 (2H,m), 2.50-2.61 (3H, m), 2.83 (1H, dd), 2.87-2.96 (2H, m), 3.04-3.21 (2H,m), 3.56-3.69 (2H, m), 3.70-3.76 (1H, m), 3.85 (3H, s), 4.30-4.37 (1H,m), 4.45(2H, dt), 4.93 (1H, br d), 5.32 (1H, s), 5.82 (1H, s), 6.76 (1H,d), 7.00 (1H, d), 7.32 (1H, s), 7.99 (1H, s), 11.00 (1H, br s). m/z: ES+[M+H]+ 497

TheN-(azetidin-3-yl)-5-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-4-methoxypyridin-2-aminewas prepared as described below

Preparation of(1S,3R)-1-(6-bromo-4-methoxypyridin-3-yl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine

6-Bromo-4-methoxynicotinaldehyde (1.46 g, 6.77 mmol) was added to asolution of(R)-3-(2-(((1-fluorocyclopropyl)methyl)amino)propyl)-2-methylaniline(0.800 g, 3.39 mmol) in AcOH (27 ml) and water (0.305 g, 16.9 mmol), andthe reaction was heated at 85° C. for 18 hours. After cooling, thereaction was concentrated under reduced pressure, and the resultingresidue was diluted with ethyl acetate and basified with saturatedaqueous sodium hydrogencarbonate. The organic layer was combined withaqueous HCl (1N), and the biphasic mixture was stirred at roomtemperature for 30 minutes. The organic layer was washed with aqueousHCl (1N), then the combined aqueous layers were extracted with ethylacetate. The aqueous layer was then basified by addition of solid K₂CO₃and extracted with ethyl acetate (2×). The combined ethyl acetateextracts from the basified aqueous layer were dried over sodium sulfate,filtered, and concentrated under reduced pressure. The resulting residuewas purified by silica flash chromatography elution gradient 10 to 100%ethyl acetate in hexanes to afford(1S,3R)-1-(6-bromo-4-methoxypyridin-3-yl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(0.450 g, 30.6%) as a gum. ¹H NMR (400 MHz, DMSO-d₆, 27° C.) δ ppm0.47-0.64 (2H, m), 0.87-0.98 (5H, m), 1.95 (3H, s), 2.42-2.48 (1H, m),2.79 (1H, dd), 2.87-2.99 (1H, m), 3.41-3.50 (H, m), 3.92-3.97 (3H, m),4.66 (2H, s), 5.11 (1H, s), 6.30 (1H, d), 6.39 (1H, d), 7.26 (1H, s),7.64 (1H, s). One hydrogen obscured by DMSO. m/z: ES+ [M+H]+ 434.

Preparation of(6S,8R)-6-(6-bromo-4-methoxypyridin-3-yl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

A solution of sodium nitrite (0.045 g, 0.65 mmol) in water (0.75 mL) wasadded dropwise to a solution of(1S,3R)-1-(6-bromo-4-methoxypyridin-3-yl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(0.270 g, 0.62 mmol) in propionic acid (3.0 mL) at −15° C. (salt/icebath), and the reaction was stirred under these conditions 1 hour.Ice-cold EtOAc (10 mL) was added followed by saturated aqueous NaHCO₃(15 mL) portionwise. The layers were separated, and the organic layerwas washed with saturated aqueous NaHCO₃ (2×). The combined aqueouslayers (pH=8) were extracted with EtOAc, and then all combined organiclayers were dried over sodium sulfate, filtered, and concentrated underreduced pressure. The resulting residue was purified by silica flashchromatography, elution gradient 20 to 60% ethyl acetate in hexanes togive(6S,8R)-6-(6-bromo-4-methoxypyridin-3-yl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.11 g, 41%) as a gum. ¹H NMR (400 MHz, DMSO-d₆, 27° C.) 0.47-0.64 (2H,m), 0.87-0.97 (2H, m), 0.97-1.01 (3H, m), 2.51-2.63 (1H, m), 2.89 (1H,dd), 2.94-3.06 (1H, m), 3.23 (1H, br dd), 3.54-3.67 (1H, m), 3.97 (3H,s), 5.28 (1H, s), 6.68 (1H, d), 7.21 (1H, d), 7.30 (1H, s), 7.68 (1H,s), 8.06 (1H, s), 12.98 (1H, br s). m/z: ES+ [M+H]+ 445.

Preparation of(6S,8R)-6-(6-bromo-4-methoxy-3-pyridyl)-7-[(1-fluorocyclopropyl)methyl]-8-methyl-3-[(2R)-tetrahydropyran-2-yl]-8,9-dihydro-6H-pyrazolo[4,3-f]isoquinolineand(6S,8R)-6-(6-bromo-4-methoxypyridin-3-yl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-2H-pyrazolo[4,3-f]isoquinoline

DCM (4 mL) was added to a flask charged with(6S,8R)-6-(6-bromo-4-methoxypyridin-3-yl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(150 mg, 0.34 mmol) and 4-methylbenzenesulfonic acid hydrate (71 mg,0.37 mmol). 3,4-Dihydro-2H-pyran (43 mg, 0.51 mmol) was added to thestirred reaction, and the reaction was stirred at RT overnight. Thereaction was washed with saturated aqueous sodium hydrogencarboante, andthe organic layer was dried over sodium sulfate, filtered, andconcentrated under reduced pressure to give crude brow gum. This waspurified by flash chromatography, elution gradient 5 to 40% EtOAc inhexanes to give a mixture of(6S,8R)-6-(6-bromo-4-methoxy-3-pyridyl)-7-[(1-fluorocyclopropyl)methyl]-8-methyl-3-[(2R)-tetrahydropyran-2-yl]-8,9-dihydro-6H-pyrazolo[4,3-f]isoquinolineand(6S,8R)-6-(6-bromo-4-methoxypyridin-3-yl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-2H-pyrazolo[4,3-f]isoquinoline(151 mg, 85%) as a gum. ¹H NMR (400 MHz, CHLOROFORM-d, 27° C.) 0.45-0.61(2H, m), 0.96-1.06 (2H, m), 1.09 (3H, d), 1.46-1.91 (3H, m), 2.03-2.20(2H, m), 2.47-2.63 (2H, m), 2.88 (1H, ddd), 3.14 (1H, dd), 3.20-3.31(1H, m), 3.64-3.79 (2H, m), 3.96 (3H, d), 3.98-4.06 (1H, m), 5.40 (1H,d), 5.64-5.71 (1H, m), 6.78 (1H, d), 6.99 (1H, d), 7.25-7.33 (1H, m),7.76 (1H, d), 8.02 (1H, d).

Preparation of tert-butyl3-((5-(((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-4-methoxypyridin-2-yl)amino)azetidine-1-carboxylateand tert-butyl3-(5-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-2H-pyrazolo[4,3-f]isoquinolin-6-yl)-4-methoxypyridin-2-ylamino)azetidine-1-carboxylate

Dioxane (2.7 mL) was added to a flask charged with a mixture of(6S,8R)-6-(6-bromo-4-methoxypyridin-3-yl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineand(6S,8R)-6-(6-bromo-4-methoxypyridin-3-yl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-2H-pyrazolo[4,3-f]isoquinoline(140 mg, 0.26 mmol), cesium carbonate (172 mg, 0.53 mmol) and tert-butyl3-aminoazetidine-1-carboxylate (68 mg, 0.40 mmol), and the reactionflask was evacuated and back-filled with nitrogen (3×). BrettPhos 3rdGeneration Precatalyst (24 mg, 0.030 mmol) was added, and the flask wasagain evacuated and back-filled with nitrogen (3×). The reaction washeated at 110° C. for 4 hours. The reaction was cooled to roomtemperature, filtered, and concentrated under reduced pressure. Theresulting um was purified by flash silica chromatography, elutiongradient 30 to 100% ethyl acetate in hexanes to give a mixture oftert-butyl3-((5-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-4-methoxypyridin-2-yl)amino)azetidine-1-carboxylateand tert-butyl3-(5-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-2H-pyrazolo[4,3-f]isoquinolin-6-yl)-4-methoxypyridin-2-ylamino)azetidine-1-carboxylate(53 mg, 32%) as a dry film. ¹H NMR (400 MHz, CHLOROFORM-d, 27° C.)0.50-0.68 (2H, m), 0.96-1.09 (2H, m), 1.12 (3H, d), 1.42-1.47 (9H, m),1.60-1.88 (3H, m), 2.07-2.21 (2H, m), 2.50-2.67 (2H, m), 2.85 (1H, ddd),3.10-3.23 (2H, m), 3.63-3.79 (4H, m), 3.87-3.91 (3H, m), 3.99-4.07 (1H,m), 4.24-4.33 (2H, m), 4.42-4.52 (1H, m), 5.36 (1H, d), 5.64-5.73 (1H,m), 5.83 (1H, d), 6.85 (1H, d), 7.26-7.32 (2H, m), 7.40 (1H, d),8.00-8.03 (1H, m). m/z: ES+ [M+H]+ 621.

Preparation ofN-(azetidin-3-yl)-5-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-4-methoxypyridin-2-amine

Methanol (0.5 mL) was added to a flask charged with tert-butyl3-((5-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-4-methoxypyridin-2-yl)amino)azetidine-1-carboxylate(0.047 g, 0.080 mmol). HCl in dioxane (4 M; 0.5 mL, 2 mmol) was added,and stirring continued for 2 hours. The reaction was then concentratedunder reduced pressure, and the resulting residue was purified using anSCX-2 cartage, eluting with 3N ammonia in methanol, to give crudeN-(azetidin-3-yl)-5-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-4-methoxypyridin-2-amine(25 mg, 76%) as a gum. The product was used in the next step withoutfurther purification. m/z: ES+ [M+H]+ 437.

Example 10N-(4-((6S,8R)-7-((3-(fluoromethyl)oxetan-3-yl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine

1-(3-Fluoropropyl)azetidin-3-amine (47.3 mg, 0.36 mmol) was added in1,4-dioxane (1.4 mL) to a sealed microwave vial containing(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-((3-(fluoromethyl)oxetan-3-yl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(100 mg, 0.18 mmol), sodium tert-butoxide (34.4 mg, 0.36 mmol) and[(2-di-cyclohexylphosphino-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl)-2-(2′-amino-1,1′-biphenyl)]palladium(II)methanesulfonate (BrettPhos Pd G3) (8.10 mg, 8.95 μmol). The vial wasdegassed with bubbling nitrogen for 10 min, then was heated to 90° C.for 1 hour. After cooling, the reaction was diluted with EtOAc andwashed with water. The aqueous phase was extracted with EtOAc, then thecombined organics were evaporated. The crude residue was dissolved inDCM (2 mL) and TFA (1 mL) was added. The mixture was stirred for 1 hour,then evaporated. The residue was dissolved in DCM and washed withsaturated aqueous NaHCO₃ solution. The aqueous layer was extracted withDCM, then the combined organics were dried over Na₂SO₄ and evaporated.The crude product was purified by preparative HPLC (Waters XBridge PrepC18 OBD column, 5μ silica, 19 mm diameter, 100 mm length), usingdecreasingly polar mixtures of water (containing 0.1% NH₃) and MeCN aseluents. Fractions containing the desired compound were evaporated todryness to affordN-(4-((6S,8R)-7-((3-(fluoromethyl)oxetan-3-yl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine(59.0 mg, 63%) as a beige solid. ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.07(3H, d), 1.68-1.87 (2H, m), 2.62 (2H, t), 2.75 (1H, d), 2.83 (1H, dd),2.87-2.96 (3H, m), 3.10 (1H, dd), 3.27 (1H, dt), 3.74 (2H, q), 3.84 (3H,s), 4.04-4.14 (2H, m), 4.43 (4H, td), 4.53 (1H, t), 4.58 (1H, d), 4.78(2H, td), 5.11 (1H, s), 5.90 (1H, dd), 6.12 (1H, d), 6.48 (1H, d), 6.80(1H, d), 7.15 (1H, d), 8.05 (1H, d); m/z: ES+[M+H]+526.

TheN-(4-((6S,8R)-7-((3-(fluoromethyl)oxetan-3-yl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-aminewas prepared as follows:

Preparation of (3-(fluoromethyl)oxetan-3-yl)methanol

Cesium fluoride (5.03 g, 33.14 mmol) was added to a flask containing(3-(bromomethyl)oxetan-3-yl)methanol (2.00 g, 11.05 mmol) in ethyleneglycol (6 mL) and the reaction was heated to 150° C. for 2 hours. Aftercooling, the reaction was diluted with EtOAc (30 mL) and water (30 mL).The aqueous was extracted with diethyl ether (3×30 mL) and ethyl acetate(3×30 mL). The combined organics were dried over MgSO₄, filtered andevaporated, then the crude product was purified by flash silicachromatography, elution gradient 0 to 100% EtOAc in heptane. Productcontaining fractions were evaporated to dryness to afford(3-(fluoromethyl)oxetan-3-yl)methanol (0.734 g, 55%) as a colourlessliquid. ¹H NMR (500 MHz, CDCl₃, 27° C.) 3.92 (2H, s), 4.52 (4H, t),4.59-4.89 (2H, m).

Preparation of (3-(fluoromethyl)oxetan-3-yl)methyltrifluoromethanesulfonate

Trifluoromethanesulfonic anhydride (1.47 ml, 8.74 mmol), followed by2,6-dimethylpyridine (1.12 ml, 9.57 mmol) were added to a solution of(3-(fluoromethyl)oxetan-3-yl)methanol (1.00 g, 8.32 mmol) in DCM (30.7mL) and the reaction was stirred at 0° C. for 1 hour. The reaction waswashed with water and 1N citric acid solution, then dried over Na₂SO₄,filtered and evaporated to afford (3-(fluoromethyl)oxetan-3-yl)methyltrifluoromethanesulfonate (1.98 g, 94%) as a red oil, which was useddirectly without further purification. ¹H NMR (500 MHz, CDCl₃, 27° C.)4.40-4.75 (8H, m).

Preparation of(R)-3-(2-(((3-(fluoromethyl)oxetan-3-yl)methyl)amino)propyl)-2-methylaniline

(3-(Fluoromethyl)oxetan-3-yl)methyl trifluoromethanesulfonate (1.92 g,7.61 mmol) was added to a solution of(R)-3-(2-aminopropyl)-2-methylaniline (1.00 g, 6.09 mmol) and DIPEA(1.58 ml, 9.13 mmol) in 1,4-dioxane (18.7 mL) and the reaction washeated to 70° C. for 5 hours. After cooling, the reaction was dilutedwith EtOAc and washed with water. The aqueous phase was extracted withEtOAc, then the combined organics were dried over Na₂SO₄, filtered andevaporated. The crude product was purified by flash silicachromatography, elution gradient 0 to 10% MeOH in EtOAc. Pure fractionswere evaporated to dryness to afford(R)-3-(2-(((3-(fluoromethyl)oxetan-3-yl)methyl)amino)propyl)-2-methylaniline(0.673 g, 42%) as a pale yellow gum. ¹H NMR (500 MHz, CDCl₃, 27° C.)1.11 (3H, d), 2.04 (3H, s), 2.59-2.71 (1H, m), 2.76-3.10 (4H, m),4.34-4.52 (4H, m), 4.53-4.68 (2H, m), 6.58 (1H, d), 6.60 (1H, d), 6.95(1H, t). m/z: ES+ [M+H]+ 267.

Preparation of(1S,3R)-1-(4-bromo-2-methoxyphenyl)-2-((3-(fluoromethyl)oxetan-3-yl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine

4-Bromo-2-methoxybenzaldehyde (645 mg, 3.00 mmol) was added to asolution of(R)-3-(2-(((3-(fluoromethyl)oxetan-3-yl)methyl)amino)propyl)-2-methylaniline(400 mg, 1.5 mmol) in acetic acid (7.4 mL) and water (135 μl, 7.50mmol). The reaction was heated to 80° C. for 4 hours. After cooling, theacetic acid was evaporated, then the residue was dissolved in DCM andwashed with saturated aqueous NaHCO₃. The organic layer was dried overNa₂SO₄, filtered and evaporated. The residue was dissolved in methanol(5 mL), then hydroxylamine hydrochloride (313 mg, 4.50 mmol) andpotassium acetate (588 mg, 6.00 mmol) were added and the reaction wasstirred at room temperature for 30 minutes. The volatiles wereevaporated, then the residue was dissolved in DCM and water. The layerswere separated, then the aqueous was extracted with DCM. The combinedorganics were dried over Na₂SO₄, filtered and evaporated, then the crudeproduct was purified by flash silica chromatography, elution gradient 0to 50% EtOAc in heptane. Product containing fractions were evaporated todryness to afford(1S,3R)-1-(4-bromo-2-methoxyphenyl)-2-((3-(fluoromethyl)oxetan-3-yl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(302 mg, 43%) as a beige solid. ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.01(3H, d), 2.07 (3H, s), 2.45 (1H, dd), 2.61-2.73 (1H, m), 2.86 (1H, d),3.01-3.14 (1H, m), 3.54 (2H, s), 3.87 (3H, s), 4.41-4.48 (2H, m), 4.51(1H, d), 4.53-4.60 (2H, m), 4.67 (1H, d), 4.73-4.81 (1H, m), 5.00 (1H,s), 6.47 (1H, s), 6.47 (1H, s), 6.61 (1H, d), 6.88 (1H, dd), 7.01 (1H,d). m/z: ES+ [M+H]+ 463.

Preparation of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-((3-(fluoromethyl)oxetan-3-yl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

Sodium nitrite (39.1 mg, 0.57 mmol) was added in water (0.5 mL) to acooled solution of(1S,3R)-1-(4-bromo-2-methoxyphenyl)-2-((3-(fluoromethyl)oxetan-3-yl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(250 mg, 0.54 mmol) in propionic acid (2.2 mL) at −15° C.(dry-ice/acetone bath). After stirring for 30 mins, ice-cold toluene (15mL) was added and the reaction was stirred at 0° C. for 15 min and thenwarmed to room temperature for 1 hour. Water (15 mL) was added and thelayers were separated. The aqueous layer was extracted with EtOAc (2×15mL), then the combined organics were washed with saturated aqueoussodium chloride, dried over Na₂SO₄, filtered and evaporated. The crudeproduct was purified by flash silica chromatography, elution gradient 0to 50% EtOAc in heptane. Product containing fractions were evaporated todryness to afford(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-((3-(fluoromethyl)oxetan-3-yl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(188 mg, 74%) as a beige solid. ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.07(3H, d), 2.70 (1H, d), 2.87 (1H, dd), 2.97 (1H, d), 3.11-3.18 (1H, m),3.18-3.27 (1H, m), 3.90 (3H, s), 4.42-4.52 (3H, m), 4.59 (1H, d),4.63-4.87 (2H, m), 5.19 (1H, s), 6.63 (1H, d), 6.76 (1H, d), 6.88 (1H,dd), 7.05 (1H, d), 7.19 (1H, d), 8.09 (1H, d). m/z: ES+ [M+H]+ 474.

Preparation of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-((3-(fluoromethyl)oxetan-3-yl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

3,4-Dihydro-2H-pyran (67.3 μl, 0.74 mmol) was added to a solution of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-((3-(fluoromethyl)oxetan-3-yl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(175 mg, 0.37 mmol) and 4-methylbenzenesulfonic acid hydrate (14.0 mg,0.07 mmol) in DCM (3.6 mL) and the reaction was stirred at 40° C. for 2hours. After cooling, the reaction was diluted with DCM and washed withsaturated aqueous NaHCO₃, dried over Na₂SO₄, filtered and evaporated.The residue was passed through a silica plug, eluting with EtOAc/heptane(1:1). The filtrate was evaporated to afford(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-((3-(fluoromethyl)oxetan-3-yl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(196 mg, 95%) as a beige solid, which was used directly in the nextstage. m/z: ES+ [M+H]+ 558.

Example 11N-(3,5-difluoro-4-((6S,8R)-7-((l-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine

4N HCl in dioxane (0.25 mL, 0.99 mmol) was added to a solution ofN-(3,5-difluoro-4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine(58 mg, 0.10 mmol) in methanol (0.30 mL) and the reaction was stirred atroom temperature for 2 hours. The volatiles were evaporated and theresidue was suspended in saturated aqueous NaHCO₃(30 mL) and extractedwith DCM (×2). The organic phase was dried over MgSO₄, filtered andevaporated. The crude residue was puried by silica gel chromatographyand eluted using a gradient of 0 to 10% 1M NH₃/MeOH in DCM. Productcontaining fractions were evaporated to dryness to affordN-(3,5-difluoro-4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine(45.4 mg, 91%) as a white solid. ¹H NMR (500 MHz, CDCl₃, 27° C.) 0.47(2H, dtd), 0.95 (2H, dt), 1.08 (3H, d), 1.68-1.85 (2H, m), 2.59 (2H, s),2.69 (1H, dd), 2.91 (3H, dq), 3.14 (1H, dd), 3.43 (1H, dd), 3.69 (2H,q), 3.81 (1H, dd), 4.01 (1H, q), 4.28 (1H, d), 4.43 (1H, t), 4.53 (1H,t), 5.19 (1H, s), 5.96 (2H, d), 6.82 (1H, d), 7.15 (1H, d), 8.06 (1H,d), 10.36 (1H, s). m/z (ES+), [M+H]+=502.

TheN-(3,5-difluoro-4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-aminewas prepared as follows:

Preparation of(1S,3R)-1-(4-bromo-2,6-difluorophenyl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine

(R)-3-(2-(((l-Fluorocyclopropyl)methyl)amino)propyl)-2-methylaniline(0.35 g, 1.48 mmol) and 4-bromo-2,6-difluorobenzaldehyde (0.33 g, 1.48mmol) in a mixture of H₂O (0.13 mL, 7.40 mmol) and acetic acid (5.79 mL)were heated at 80° C. for 7 hours. The solvent was evaporated and theresidue was treated with HCl (1N, 10 mL) and was stirred at roomtemperature for 1 hour. To the reaction was added solid Na₂CO₃ until thereaction pH was >8. The mixture was diluted with H₂O (40 mL) andextracted twice with EtOAc (2×50 mL). The organic phase was dried overMgSO₄, filtered, concentrated and was purified by silica chromatography(10 to 70% EtOAc in heptanes) to afford(1S,3R)-1-(4-bromo-2,6-difluorophenyl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(0.285 g, 44%) as a pale yellow gum. ¹H NMR (500 MHz, CDCl₃, 27° C.)0.44 (2H, dq), 0.91-0.95 (1H, m), 0.95-0.99 (1H, m), 1.02 (3H, d), 2.06(3H, s), 2.50-2.65 (2H, m), 3.00-3.15 (2H, m), 3.50 (2H, s), 3.69 (1H,d), 5.17 (1H, s), 6.42 (2H, s), 6.97 (1H, d), 6.99 (1H, d). m/z (ES+),[M+H]+=439.

Preparation of(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

(1S,3R)-1-(4-Bromo-2,6-difluorophenyl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(0.280 g, 0.64 mmol) in propionic acid (2.66 mL) was cooled to −17° C.(dry ice/acetone bath). Sodium nitrite (0.044 g, 0.64 mmol) in water(0.53 mL) was added dropwise and the reaction mixture was stirred at−17° C. for 30 minutes. The reaction mixture was diluted with ice-coldtoluene (15 mL) and stirred at 4° C. for 15 minutes. The mixture wasthen stirred and warmed to room temperature for 45 minutes. The reactionmixture was washed with water (2×15 mL), the combined aqueous phaseswere washed with EtOAc (2×10 mL), the combined organics washed withsaturated aqueous sodium chloride (1×20 mL), dried (MgSO₄), filtered andthe filtrate evaporated to an orange-brown oil. The crude material waspurified by flash silica chromatography, elution gradient 0-40% EtOAc inheptane to afford(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.216 g, 75%) as an off-white solid. ¹H NMR (500 MHz, CDCl₃, 27° C.)0.46 (2H, dddd), 0.91-1.04 (2H, m), 1.07 (3H, d), 2.66 (1H, dd), 2.95(1H, dd), 3.15 (1H, dd), 3.46 (1H, dd), 3.83 (1H, dd), 5.32 (1H, s),6.76 (1H, d), 7.00 (2H, d), 7.18 (1H, d), 8.09 (1H, d), 10.52 (1H, s).m/z (ES+), [M+H]+=450.

Preparation of(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineand(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

A microwave vial was charged with(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(216 mg, 0.48 mmol), 4-methylbenzenesulfonic acid hydrate (9.1 mg, 0.05mmol), 3,4-dihydro-2H-pyran (0.07 mL, 0.72 mmol) and DCM (2 mL). Themixture was heated in a microwave at 80° C. for 20 minutes. A further(0.035 mL, 0.36 mmol) of 3,4-dihydro-2H-pyran was added and the reactionwas heated to 85° C. for a further 15 minutes. The reaction was dilutedwith DCM and washed with saturated aqueous NaHCO₃, then the organiclayer was dried over Na₂SO₄, filtered and concentrated. The crudeproduct was purified by flash silica chromatography, elution gradient 0to 50% EtOAc in heptane. Product containing fractions were evaporated todryness to afford two regioisomeric THP-protected products(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineand(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(237 mg, 92%) as a pale yellow gum (data for major isomer given). ¹H NMR(500 MHz, CDCl₃, 27° C.) 0.35-0.55 (2H, m), 0.96 (2H, ddd), 1.05 (3H,dd), 1.58-1.84 (3H, m), 1.97-2.32 (3H, m), 2.64 (1H, dd), 2.77 (1H, dd),3.14 (1H, ddd), 3.29-3.54 (1H, m), 3.62-3.91 (2H, m), 3.96-4.25 (1H, m),5.26 (1H, d), 5.65 (1H, ddd), 6.64 (1H, d), 7.00 (2H, d), 7.26-7.45 (1H,m), 8.13 (1H, dd). m/z (ES+), [M+H]+=534.

Preparation ofN-(3,5-difluoro-4-((6S,8R)-7-((l-fluorocyclopropyl)methyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine

[(2-Di-cyclohexylphosphino-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl)-2-(2′-amino-1,1′-biphenyl)]palladium(II)methanesulfonate (BrettPhos Pd G3) (5.0 mg, 5.54 μmol) was added to asuspension of(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineand(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(37 mg, 0.07 mmol), 1-(3-fluoropropyl)azetidin-3-amine (22.9 mg, 0.17mmol) and sodium tert-butoxide (13.3 mg, 0.14 mmol) in degassed1,4-dioxane (0.58 mL) and the reaction was heated to 95° C. for 1.5hours in the microwave. The reaction mixture was diluted with EtOAc,filtered through celite and evaporated to dryness. The crude product waspurified by flash silica chromatography, elution gradient 0 to 10% 1MNH₃/MeOH in DCM. Pure fractions were evaporated to dryness to affordN-(3,5-difluoro-4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine(18.0 mg, 44%) as a colourless dry film. ¹H NMR (500 MHz, CDCl₃, 27° C.)0.38-0.54 (2H, m), 0.94 (2H, ddd), 1.06 (3H, dd), 1.67-1.82 (4H, m),2.04-2.25 (4H, m), 2.59 (2H, t), 2.63-2.77 (2H, m), 2.89 (2H, dd), 3.14(1H, s), 3.29-3.37 (1H, m), 3.70 (2H, q), 3.75-3.81 (2H, m), 4.00 (1H,q), 4.12-4.17 (1H, m), 4.22 (1H, d), 4.43 (1H, t), 4.52 (1H, t), 5.10(1H, s), 5.55-5.74 (1H, m), 5.96 (2H, d), 6.71 (1H, d), 7.28-7.44 (1H,m), 8.10 (1H, dd). m/z (ES+), [M+H]+=586.

Examples 12 & 13 Preparation of individual diastereoisomers of(6S,8R)-7-(2-fluoro-3-methoxy-2-methylpropyl)-6-(4-(1-(3-fluoropropyl)azetidin-3-yloxy)-2-methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

DMF (0.66 mL) followed by DIPEA (23.9 μl, 0.14 mmol) were added to aflask charged with(6S,8R)-6-(4-(azetidin-3-yloxy)-2-methoxyphenyl)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(32 mg, 0.07 mmol). Into the stirred reaction was injected dropwise1-fluoro-3-iodopropane (13.5 mg, 0.07 mmol) and the reaction was stirredat room temperature overnight. The reaction was evaporated to drynessand the residue was purified by flash silica chromatography, elutiongradient 0 to 10% 1M NH₃/MeOH in DCM. Product containing fractions wereevaporated to dryness to afford the product as a diastereoisomericmixture. The diastereoisomers were separated by chiral preparative SFC(Phenomonex Lux C4 column, 5μ silica, 30 mm diameter, 250 mm length),using decreasingly isocratic 40% MeOH+0.1% NH₃ in CO₂ as eluent toafford the first eluting isomer (8.0 mg, 22%) and the second elutingisomer (8.0 mg, 22%) as pale yellow dry films.

Isomer 1: ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.06 (3H, d), 1.28 (3H, d),1.72-1.83 (2H, m), 2.47 (1H, dd), 2.63 (2H, t), 2.8-2.95 (2H, m),3.03-3.11 (2H, m), 3.19 (2H, s), 3.33 (3H, s), 3.56 (1H, dd), 3.67 (1H,d), 3.79 (2H, q), 3.86 (3H, s), 4.44 (1H, t), 4.53 (1H, t), 4.74 (1H,t), 5.32 (1H, s), 6.10 (1H, dd), 6.41 (1H, d), 6.69 (1H, d), 6.76 (1H,d), 7.13 (1H, d), 8.05 (1H, d), 10.20 (1H, s). m/z (ES+), [M+H]+=529;

Isomer 2: ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.06 (3H, d), 1.25 (3H, d),1.70-1.82 (2H, m), 2.45-2.56 (1H, m), 2.63 (2H, t), 2.78-2.91 (2H, m),3.02-3.11 (2H, m), 3.18 (1H, d), 3.36 (3H, s), 3.42 (1H, dd), 3.47-3.55(1H, m), 3.62 (1H, d), 3.74-3.82 (2H, m), 3.86 (3H, s), 4.43 (1H, t),4.53 (1H, t), 4.73 (1H, t), 5.36 (1H, s), 6.08 (1H, dd), 6.41 (1H, d),6.67 (1H, d), 6.79 (1H, d), 7.15 (1H, d), 8.06 (1H, d), 10.20 (1H, s).m/z (ES+), [M+H]+=529.

The(6S,8R)-6-(4-(azetidin-3-yloxy)-2-methoxyphenyl)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolinewas prepared as follows:

Preparation of a diastereoisomeric mixture of(6S,8R)-6-(4-(azetidin-3-yloxy)-2-methoxyphenyl)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

Tert-butyl 3-hydroxyazetidine-1-carboxylate (102 mg, 0.59 mmol),(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(110 mg, 0.20 mmol), tert-butyl 3-hydroxyazetidine-1-carboxylate (102mg, 0.59 mmol), Pd RockPhos 3rd generation (16.6 mg, 0.02 mmol) andcesium carbonate (128 mg, 0.39 mmol) were suspended in toluene (1.26 mL)and sealed into a microwave tube. The reaction was heated to 95° C. for20 hours. The reaction mixture was diluted with EtOAc (25 mL) andfiltered through celite. The crude product was purified by flash silicachromatography, elution gradient 0 to 100% EtOAc in heptane. Thefractions containing the impure product were evaporated to dryness anddissolved in DCM (1.3 mL) to which TFA (378 μl, 4.91 mmol) was addeddropwise. The reaction was stirred at room temperature for 2 hours, thenvolatiles were removed in vacuo. The residue was washed with saturatedaqueous NaHCO₃ (30 mL) and extracted with DCM (2×30 mL). The combinedorganic phase was dried over MgSO₄, filtered and evaporated. The crudeproduct was purified by flash silica chromatography, elution gradient 0to 40% 1M NH₃/MeOH in DCM. Pure fractions were evaporated to dryness toafford(6S,8R)-6-(4-(azetidin-3-yloxy)-2-methoxyphenyl)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(35.0 mg, 38%) as a white solid. ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.06(3H, dd), 1.24-1.35 (3H, m), 2.49 (1H, dt), 2.76-2.96 (2H, m), 3.08-3.27(2H, m), 3.34 (3H, d), 3.41 (1H, s), 3.53-3.69 (2H, m), 3.69-3.93 (7H,m), 4.81-5.08 (1H, m), 5.34 (1H, d), 6.02-6.12 (1H, m), 6.40 (1H, t),6.68 (1H, s), 6.77 (1H, dd), 7.13 (1H, d), 8.05 (1H, t). m/z (ES+),[M+H]+=469.

Examples 14 & 15 Preparation of individual diastereoisomers ofN-(4-((6S,8R)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine

1-(3-Fluoropropyl)azetidin-3-amine (175 mg, 1.32 mmol),(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(370 mg, 0.66 mmol) and sodium tert-butoxide (127 mg, 1.32 mmol) weresuspended in 1,4-dioxane (8 mL). The mixture was degassed and[(2-di-cyclohexylphosphino-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl)-2-(2′-amino-1,1′-biphenyl)]palladium(II)methanesulfonate methanesulfonate (Brett Phos G3) (60 mg, 0.07 mmol) wasadded. The reaction was heated to 100° C. for 3 hours. The reactionmixture was diluted with DCM (100 mL) and washed with water (100 mL).The organic layer was evaporated. This process was repeated using 114 mg(0.3 mmol) of the aryl bromide. The combined crude products from thesereactions was suspended in hydrochloric acid (2M, 5 mL) and stirred atroom temperature for 2 hours. The reaction mixture was diluted withwater (10 mL) and washed with EtOAc (10 mL). The aqueous phase wasbasified with 2M aqueous sodium hydroxide and extracted with DCM (2×15mL). The combined organics were evaporated and the crude product waspurified by flash silica chromatography, elution gradient 0 to 20% MeOHin DCM. Fractions were evaporated to dryness to afford the crude productas a mixture of diastereoisomers which were separated by chiralpreparative SFC (Phenomonex Lux C4 column, 5μ silica, 30 mm diameter,250 mm length), using decreasingly isocratic 40% MeOH+0.1% NH₃ in CO₂ aseluent to afford the first eluting isomer (42 mg, 19%) and the secondeluting isomer (29 mg, 13%)

Isomer 1: ¹H NMR (500 MHz, DMSO, 27° C.) 0.96 (3H, d), 1.20 (3H, d),2.36-2.44 (1H, m), 2.65-2.85 (3H, m), 2.92 (4H, q), 3.05-3.22 (3H, m),3.23 (3H, s), 3.24-3.27 (2H, m), 3.46-3.59 (2H, m), 3.78 (3H, s), 4.49(2H, dd), 5.16 (1H, s), 5.35 (1H, t), 5.95 (1H, dd), 6.23 (1H, d), 6.36(1H, d), 6.63 (1H, d), 7.16 (1H, d), 8.02 (1H, s), 12.90 (1H, s); ¹⁹FNMR (471 MHz, DMSO, 27° C.) −219.77, −149.11, m/z: ES+ [M+H]+ 528.

Isomer 2: ¹H NMR (500 MHz, DMSO, 27° C.) 0.97 (3H, d), 1.19 (3H, d),2.68-2.81 (3H, m), 2.88-2.95 (3H, m), 3.05-3.11 (1H, m), 3.21-3.28 (5H,m), 3.32-3.39 (1H, m), 3.43-3.56 (2H, m), 3.79 (3H, s), 4.49 (2H, dd),5.17 (1H, s), 5.34 (1H, t), 5.93 (1H, dd), 6.23 (1H, d), 6.35 (1H, d),6.64 (1H, d), 7.16 (1H, d), 8.02 (1H, s), 12.90 (1H, s); ¹⁹F NMR (471MHz, DMSO, 27° C.) −219.76, −148.20; m/z: ES+ [M+H]+ 528.

Example 16 Preparation of2,2-difluoro-34(6S,8R)-6-(54(1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol

Hydrochloric acid (2.0 M, 0.5 mL) was added to2,2-difluoro-3-((6S,8R)-6-(5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol(0.132 g, 0.23 mmol) and the solution was stirred at room temperaturefor 3 hours. The reaction mixture was purified by ion exchangechromatography, using an SCX column. The desired product was eluted fromthe column using 1M NH₃/MeOH to afford2,2-difluoro-3-((6S,8R)-6-(5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol(0.084 g, 75%) as a white solid. ¹H NMR (500 MHz, DMSO, 27° C.) 1.04(3H, d), 1.20-1.28 (2H, m) 1.59-1.71 (2H, m), 2.60-2.70 (1H, m),2.73-2.84 (3H, m), 2.99 (1H, dd), 3.08-3.18 (1H, m), 3.41 (1H, d),3.61-3.72 (4H, m), 3.93 (1H, d), 4.44 (2H, dt), 4.94 (1H, s), 5.42 (1H,t), 6.21 (1H, d), 6.78-6.82 (2H, m), 6.90 (1H, d), 7.21 (1H, d), 7.74(1H, d), 8.03 (1H, s), 12.94 (1H, s); ¹H NMR (471 MHz, DMSO, 27° C.)−218.24, −108.31; m/z: ES+ [M+H]+ 489.

The2,2-difluoro-3-((6S,8R)-6-(5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-olwas prepared as follows:

Preparation of(1S,3R)-1-(5-bromopyridin-2-yl)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine

(R)-3-(2-((3-((Tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)amino)propyl)-2-methylaniline(1.20 g, 2.42 mmol) and 5-bromopicolinaldehyde (0.944 g, 5.07 mmol) wereheated in acetic acid (12 mL) and water (0.22 mL, 12.08 mmol) to 70° C.for 30 minutes. The reaction mixture was evaporated and the residue wasredisolved in ethanol (10 mL). Potassium acetate (0.594 g, 6.05 mmol)and hydroxylamine hydrochloride (0.252 g, 3.63 mmol) were added. Thereaction mixture was stirred at room temperature for 30 minutes thenevaporated and the residue partitioned between DCM (30 mL) and saturatedaqueous sodium bicarbonate (30 mL). The organic phase was evaporated andthe crude product was purified by flash silica chromatography, elutiongradient 0 to 50% EtOAc in heptane to afford(1S,3R)-1-(5-bromopyridin-2-yl)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(1.090 g, 68%) as a colourless gum. ¹H NMR (500 MHz, DMSO, 27° C.) 0.96(9H, s), 0.98 (3H, d), 1.94 (3H, s), 2.40 (1H, dd), 2.57 (1H, dd),2.66-2.78 (1H, m), 3.11-3.22 (2H, m), 3.84-3.93 (1H, m), 3.95-4.05 (1H,m), 4.69 (2H, s), 4.91 (1H, s), 6.39 (1H, d), 6.42 (1H, d), 6.99 (1H,d), 7.39-7.48 (6H, m), 7.59 (4H, ddd), 7.80 (1H, dd), 8.51 (1H, dd); ¹HNMR (471 MHz, DMSO, 27° C.) −109.84 (d), −108.39 (d); m/z: ES+ [M+H]+664/666.

Preparation of(6S,8R)-6-(5-bromopyridin-2-yl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

Sodium nitrite (119 mg, 1.72 mmol) in water (1.0 mL) was added to an icecooled solution of(1S,3R)-1-(5-bromopyridin-2-yl)-2-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(1.09 g, 1.64 mmol) and para-toluenesulfonic acid hydrate (0.94 g, 4.92mmol) in acetonitrile (6 mL). The reaction was stirred for 15 minutesand then warmed to room temperature for 45 minutes. After cooling backto 4° C., tetrabutylammonium acetate (2.54 g, 8.20 mmol) in ice-coldMeCN (40 mL) was added and the reaction was stirred for 15 minutesbefore warming to room temperature for a further 30 minutes, EtOAc (120mL) was added followed by 2 M NaOH (100 mL). The organic phase waswashed with saturated aqueous sodium chloride, dried (Na₂SO₄) andevaporated. The crude product was purified by flash silicachromatography, elution gradient 0 to 50% EtOAc in heptane to afford(6S,8R)-6-(5-bromopyridin-2-yl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.330 g, 30%) as a light brown foam.

¹H NMR (500 MHz, DMSO, 27° C.) 0.97 (9H, s), 1.06 (3H, d), 2.75-2.91(2H, m), 2.99-3.06 (1H, m), 3.34-3.41 (2H, m), 3.88-4.01 (2H, m), 5.12(1H, s), 6.83 (1H, d), 7.12 (1H, d), 7.26 (1H, d), 7.39-7.45 (4H, m),7.46-7.50 (2H, m), 7.56-7.62 (4H, m), 7.85 (1H, dd), 8.07-8.09 (1H, m),8.54 (1H, dd), 13.02 (1H, s); ¹⁹F NMR (471 MHz, DMSO, 27° C.) −110.01(d), −108.72 (d); m/z: ES+ [M+H]+ 675/677.

Preparation of a diastereoisomeric mixture of(6S,8R)-6-(5-bromopyridin-2-yl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

To a solution of(6S,8R)-6-(5-bromopyridin-2-yl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.330 g, 0.49 mmol) and 3,4-dihydro-2H-pyran (0.13 mL, 1.47 mmol) inDCM (5 mL) was added para toluenesulfonic acid hydrate (93 mg, 0.49mmol) and the reaction heated at 40° C. for 1 hour. The reaction mixturewas diluted with DCM (20 mL) and washed with saturated aqueous sodiumbicarbarbonate (20 mL). The organic phase was evaporated to a dark brownoil and the crude product was purified by flash silica chromatography,elution gradient 0 to 50% EtOAc in heptane to afford(6S,8R)-6-(5-bromopyridin-2-yl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-3-((S)-tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.330 g, 89%) as a gum. ¹⁹F NMR (471 MHz, DMSO, 27° C.) −110.03 (d),−108.83 (d), −108.79 (d); m/z: ES+ [M+H]+ 759/761.

Preparation of a diastereoisomeric mixture of6-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine

1-(3-Fluoropropyl)azetidin-3-amine (0.115 g, 0.87 mmol),(6S,8R)-6-(5-bromopyridin-2-yl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.330 g, 0.43 mmol),dicyclohexyl(2′,4′,6′-triisopropyl-3,6-dimethoxy-[1,1′-biphenyl]-2-yl)phosphane(0.023 g, 0.04 mmol) and sodium tert-butoxide (0.125 g, 1.30 mmol) weresuspended in 1,4-dioxane (4 mL). The mixture was degassed and[(2-di-cyclohexylphosphino-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl)-2-(2′-amino-1,1′-biphenyl)]palladium(II)methanesulfonate methanesulfonate (BrettPhos G3) (0.039 g, 0.04 mmol)was added. The reaction was heated to 100° C. for 3 hours. The reactionmixture was diluted with DCM (50 mL) and washed with water (50 mL). Theorganic layer was evaporated and the crude product was purified by flashsilica chromatography, elution gradient 0 to 20% MeOH in DCM to afford6-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine(0.264 g, 75%) as a yellow gum as a mixture of diastereoisomers. ¹⁹F NMR(471 MHz, DMSO, 27° C.) −218.19, −110.13, −109.44, −108.6 −107.8; m/z:ES+ [M+H]+ 811.

Preparation of a diastereoisomeric mixture of2,2-difluoro-3-((6S,8R)-6-(5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol

Tetrabutylammonium fluoride (1.0 M in THF, 0.5 mL, 0.50 mmol) was addedto solution of6-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine(0.264 g, 0.33 mmol) in THF (2 mL). The reaction mixture was stirred atroom temperature for 4 hours and the reaction mixture was evaporated andthe residue partitioned between DCM (20 mL) and water (20 mL). Theorganic phase was evaporated and the crude product was purified by flashsilica chromatography, elution gradient 0 to 20% MeOH in DCM to afford2,2-difluoro-3-((6S,8R)-6-(5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol(0.132 g, 71%) as a colourless dry film as a mixture ofdiastereoisomers. ¹⁹F NMR (471 MHz, DMSO, 27° C.) −218.19, −108.42,−108.39; m/z: ES+ [M+H]+ 573.

Example 17 Preparation ofN-(1-(3-fluoropropyl)azetidin-3-yl)-64(6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine

1-(3-Fluoropropyl)azetidin-3-amine (50 mg, 0.38 mmol),(6S,8R)-6-(5-bromopyridin-2-yl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.100 g, 0.20 mmol), cesium carbonate (128 mg, 0.39 mmol) and[(2-di-cyclohexylphosphino-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl)-2-(2′-amino-1,1′-biphenyl)]palladium(II)methanesulfonate methanesulfonate (Brett Phos G3) (23 mg, 0.02 mmol)were suspended in 1,4-dioxane (2 mL) and sealed into a microwave tube.The reaction was heated to 100° C. for 4 hours under microwaveirradiation. The reaction mixture was diluted with DCM (20 mL) andwashed with water (20 mL). The organic layer was evaporated and thecrude product was dissolved in DCM (3 mL) and TFA (0.3 mL). The reactionwas stirred at room temperature for 1 hour then the reaction mixture waspartitioned between DCM (20 mL) and 2M NaOH (20 mL). The organic phasewas evaporated and the crude product was purified by flash silicachromatography, elution gradient 0 to 20% MeOH in DCM. Productcontaining fractions were evaporated to dryness to affordN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine(9.0 mg, 10%) as a colourless dry film. ¹H NMR (500 MHz, CDCl₃, 27° C.)1.14 (3H, d), 1.70-1.82 (2H, m), 2.62 (2H, t), 2.85 (1H, dd), 2.93-3.06(3H, m), 3.20-3.31 (2H, m), 3.59 (1H, td), 3.73 (2H, dt), 4.10 (2H, dd),4.48 (2H, dt), 5.03 (1H, s), 6.80 (1H, dd), 6.89 (1H, d), 7.15 (1H, d),7.21 (1H, d), 7.84 (1H, dd), 8.01 (1H, d); ¹⁹F NMR (471 MHz, DMSO, 27°C.) −224.69, −75.55; m/z: ES+ [M+H]+ 477.

The(6S,8R)-6-(5-bromopyridin-2-yl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolinewas prepared as follows.

Preparation of(1S,3R)-1-(5-bromopyridin-2-yl)-3,5-dimethyl-2-(2,2,2-trifluoroethyl)-1,2,3,4-tetrahydroisoquinolin-6-amine

(R)-2-Methyl-3-(2-((2,2,2-trifluoroethyl)amino)propyl)aniline (0.290 g,1.18 mmol) and 5-bromopicolinaldehyde (0.460 g, 2.47 mmol) were heatedin acetic acid (6 mL) and water (0.1 mL) to 70° C. for 30 minutes. Thereaction mixture was evaporated and the residue dissolved in ethanol (10mL). Potassium acetate (0.290 g, 2.95 mmol) and hydroxylaminehydrochloride (0.123 g, 1.77 mmol) were added and the reaction mixturewas stirred at room temperature for 30 minutes. The reaction mixture wasevaporated and the residue was partitioned between DCM (30 mL) andsaturated aqueous sodium bicarbonate (30 mL). The organic phase wasevaporated and the crude product was purified by flash silicachromatography, elution gradient 0 to 50% EtOAc in heptane to afford(1S,3R)-1-(5-bromopyridin-2-yl)-3,5-dimethyl-2-(2,2,2-trifluoroethyl)-1,2,3,4-tetrahydroisoquinolin-6-amine(0.264 g, 54%) as a colourless dry film. ¹H NMR (500 MHz, DMSO, 27° C.)1.03 (3H, d), 1.94 (3H, s), 2.45 (1H, dd), 2.67 (1H, dd), 2.90 (1H, dd),3.22-3.29 (1H, m), 3.49 (1H, dd), 4.69 (2H, s), 4.85 (1H, s), 6.40-6.46(2H, m), 7.21 (1H, d), 7.95 (1H, dd), 8.56 (1H, dd); ¹⁹F NMR (471 MHz,DMSO, 27° C.) −69.83; m/z: ES+ [M+H]+ 414/416.

Preparation of(6S,8R)-6-(5-bromopyridin-2-yl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

Sodium nitrite (0.048 g, 0.70 mmol) was added in water (0.2 mL) to acooled solution of(1S,3R)-1-(5-bromopyridin-2-yl)-3,5-dimethyl-2-(2,2,2-trifluoroethyl)-1,2,3,4-tetrahydroisoquinolin-6-amine(0.264 g, 0.64 mmol) in propionic acid (2.5 mL) at −10° C. The reactionwas stirred for 1 h, then ice-cold EtOAc (10 mL) was added. The reactionwas quenched by addition of aqueous saturated sodium bicarbonate (15 mL)and stirred for 15 minutes before being allowed to warm to roomtemperature. The organic phase was dried (Na₂SO₄) and evaporated to givecrude(6S,8R)-6-(5-bromopyridin-2-yl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.272 g, 0.64 mmol) as a brown gum. The material was dissolved in DCM(10 mL) and para toluenesulfonic acid hydrate (0.012 g, 0.06 mmol) wasadded and the mixture heated at 40° C. for 2 hours. The reaction mixturewas diluted with DCM (20 mL) and washed with aqueous NaOH (2M, 30 mL).The organic phase was evaporated to a dark brown oil and the crudeproduct was purified by flash silica chromatography, elution gradient 0to 40% EtOAc in heptane to afford(6S,8R)-6-(5-bromopyridin-2-yl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,34]isoquinoline(0.142 g, 44%). m/z: ES+ [M+H]+ 509/511.

Alternative route #1 to Example 17: Preparation ofN-[1-(3-fluoropropyl)azetidin-3-yl]-6-[(6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-3,6,8,9-tetrahydropyrazolo[4,3-f]isoquinolin-6-yl]pyridin-3-amine

Trifluoro acetic acid (1.30 L, 17.04 mol) was added to a stirredsolution of tert-butyl (1-(3-fluoropropyl)azetidin-3-yl)carbamate (0.50Kg, 2.04 mol) in DCM (1.50 L) at 0° C. over 0.5 hours. The mixture wasallowed to warm to 20° C. and stirred at 20° C. for 2.5 hours. Themixture was heated to 30° C. and stirring was continued for 1 hour. Themixture was concentrated under reduced pressure and the residue wasconcentrated under reduced pressure from toluene (3×3.00 L). Theresulting residue was dissolved in 1,4-dioxane (3.00 L) and treated with30% sodium 2-methylbutan-2-olate solution in Me-THF (5.50 L, 13.63 mol)over 45 minutes, ensuring the temperature did not exceed 29° C. Asolution of 90% w/w(6S,8R)-6-(5-bromopyridin-2-yl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.81 Kg, 1.70 mol) in 1,4-dioxane (1.50 L) and BrettPhos Pd G3 (0.077Kg, 0.085 mol) were added and the atmosphere was replaced with nitrogenusing cycles of vacuum and nitrogen (3×). The mixture was heated to 55°C. and stirred at 55° C. 2.5 hours. The mixture was cooled to 21° C. andan 8% NaHCO₃ solution in water (7.16 L, 6.82 mol) was added, followed byisopropyl acetate (4.50 L). The mixture was stirred for 2-3 minutes andthe layers were separated. The organic layer was washed with 8% NaHCO₃solution in water (7.16 L, 6.82 mol), stirring the mixture for 5 minutesand then the layers were separated. The combined aqueous layers wereextracted with isopropyl acetate (4.50 L). The combined organic layerswere treated with Silicycle (SiliaMetS-Thiol, 400 g, 1.26 mmol/g, 6 eqvs. Pd) and the mixture was stirred for 16 hours. The solids wereremoved by filtration through celite, washing the filter-cake withisopropyl acetate. The filtrate was concentrated under reduced pressure(bath temperature 40° C.). The residue was dissolved in ispropyl acetate(5.00 L) and treated with further Silicycle (SiliaMetS-Thiol, 250 g,1.26 mmol/g, 3.7 eq vs. Pd). The resulting mixture was stirred at 21° C.for 16 hours. The solids were removed by filtration through celite,washing the filter-cake with isopropyl acetate (2.50 L). The filtratewas concentrated under reduced pressure (bath temperature 40° C.) togive a brown foam. The crude product was purified by preparative SFC(Lux C4 column, 50 mm diameter, 250 mm length), using a gradient elutionof a EtOH/DEA 100/0.5 in CO₂, 140 bar mobile phase at a flow rate of 400mL/minute at 40° C.) to give a light brown solid foam. EtOAc (2.00 L)was added and the solution was concentrated under reduced pressure.EtOAc (1.90 L) and heptane (1.90 L) were added and the mixture wasseeded at 25° C. The mixture was kept at 25° C. for 1 hour. Heptane(6.50 L) was added over 2 hours to the stirring mixture. The resultingmixture was stirred at 22° C. for 18 hours. The resulting solid wascollected by filtration, washing the filter-cake with 1:5 EtOAc/heptane(2.50 L). The solid was sucked dry under a nitrogen blanket for 30minutes, then dried under vacuum at 40° C. for 9 days to giveN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine(626 g, 77%) as an off-white solid. ¹H NMR (600 MHz, DMSO-d₆, 27° C.)1.07 (3H d) 1.63 (2H, dquin), 2.45 (2H, t), 2.72 (2H, br t), 2.83 (1H,br dd), 2.91-3.00 (1H, m), 3.06 (1H, br dd) 3.42-3.55 (2H, m), 3.58-3.65(2H, m), 3.92 (1H, dquin) 4.43 (2H, dt) 4.92 (1H, s), 6.22 (1H, d), 6.79(1H, d) 6.82 (1H, dd), 6.96 (1H, d), 7.21 (1H, d), 7.73 (1H, d), 8.04(1H, s), 12.96 (1H, s). m/z: ES+ [M+H]+ 477.

Procedures used to prepare the starting materials tert-butyl(1-(3-fluoropropyl)azetidin-3-yl)carbamate and(6S,8R)-6-(5-bromopyridin-2-yl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineare described below.

Preparation of tert-butyl (1-(3-fluoropropyl)azetidin-3-yl)carbamate

Reaction was performed in duplicate and combined for workup:1-Fluoro-3-iodo-propane (189 g, 1.00 mol) was added to a stirred mixtureof tert-butyl N-(azetidin-3-yl)carbamate;hydrochloride (200 g, 0.96 mol)and K₂CO₃ (331 g, 2.40 mol) in THF (1.40 L) and water (0.50 mol) at 20°C. The reaction mixture was heated to 70° C. and stirred at 70° C. for1.5 hours. The two reaction mixtures combined and added to water (2.5 L)and EtOAc (2.0 L). The mixture was stirred at 21° C. for 3 minutes. Thelayers were separated and the organic layer was washed with water (2×2.0L) and saturated ammonium chloride solution (2.0 L). The organic layerwas concentrated under reduced pressure to give the crude product.Toluene (400 mL) was added and the mixture was heated to 60° C. Heptane(1.6 L) was added and the mixture was allowed to cool to 21° C. andstirred for 15 minutes. The solid was collected by filtration and suckeddry to give tert-butyl (1-(3-fluoropropyl)azetidin-3-yl)carbamate (230g, 52%) as a white solid. ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.44 (9H, s),1.68-1.80 (2H, m), 2.56 (2H, t), 2.87 (2H, s), 3.66 (2H, t), 4.29 (1H,s), 4.43 (1H, t), 4.52 (1H, t), 4.86 (1H, s).

Preparation of 4-bromo-1-tetrahydropyran-2-yl-indazole

To a slurry of 4-bromo-1H-indazole (1.50 Kg, 7.60 mol) and3,4-dihydro-2H-pyran (0.96 Kg, 11.40 mol) in DCM (2.40 L) was added4-methylbenzenesulfonic acid hydrate (11.60 g, 0.06 mol) at 20° C. undernitrogen. The resulting slurry was stirred at between 20° C. and 29° C.(small exotherm) for 110 minutes. The resulting solution was washed withsaturated aqueous NaHCO₃, (2.00 L), and the organic layer was evaporatedunder reduced pressure. Hot (70° C.) heptane (2.50 L) was added and themixture was evaporated under reduced pressure to give a crystallinesolid. Purification by recrystallisation from hot (70° C.) heptane (7.00L) was carried out allowing the solution to cool slowly to 34° C. Theresulting solid was collected by filtration and washed with cold heptaneto give 4-bromo-1-tetrahydropyran-2-yl-indazole (1.87 Kg, 87%) as acrystalline solid. ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 1.54-1.66 (2H, m),1.67-1.85 (1H, m), 1.94-2.09 (2H, m), 2.31-2.45 (1H, m), 3.70-3.81 (1H,m), 3.83-3.97 (1H, m), 5.88 (1H, dd), 7.32-7.39 (1H, m), 7.40-7.44 (1H,m), 7.79 (1H, dt), 8.09 (1H, d) m/z: ES− [M−H]⁻ 282.

Preparation of tert-butylN-[(1R)-1-methyl-2-(1-tetrahydropyran-2-ylindazol-4-yl)ethyl]carbamate

A solution of 4-bromo-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole (0.70 Kg,2.49 mol) in THF (3.50 L) was gradually cooled in 10° C. increments to−70° C. 2.5 M Butyllithium in hexanes (1.10 L, 2.74 mol) was added over70 minutes, keeping the internal temperature below −60° C. The resultingmixture was stirred at −72° C. for 1 hour. 2.3 M hexyllithium in hexanes(0.054 L, 0.12 mol) was added, followed by a solution of tert-butyl(R)-4-methyl-1,2,3-oxathiazolidine-3-carboxylate 2,2-dioxide (652 g,2.74 mol) in THF (2.80 L) over 2 hours, ensuring the internaltemperature did not exceed −58° C. The resulting mixture was stirred at−60° C. for 16 hours. The mixture was slowly warmed to −10° C. over 1.5hours. Water (2.10 L) was carefully added. The layers were separated andthe organic layer was washed with saturated brine (1.40 L) and theorganic layer was concentrated under reduced pressure to give an oil.IPA (1.50 L) was added and the mixture was concentrated under reducedpressure to give tert-butyl((2R)-1-(1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)propan-2-yl)carbamate(1.27 Kg, >100%) as a brown oil. This was taken on without furtherpurification. ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.08 (3H, dd), 1.43 (9H,s), 1.63-1.69 (1H, m), 1.73-1.82 (2H, m), 2.07 (1H, dd), 2.17 (1H, dd),2.53-2.64 (1H, m), 2.95 (1H, s), 3.21 (1H, dt), 3.70-3.80 (1H, m),3.97-4.12 (2H, m), 4.33-4.49 (1H, m), 5.71 (1H, ddd), 6.96 (1H, dd),7.31 (1H, ddd), 7.46 (1H, d), 8.13 (1H, s). m/z: ES+ [M.+] 359

Preparation of (2R)-1-(1H-indazol-4-yl)propan-2-amine.dihydrochloride

12 M HCl (1.00 L, 12.0 mol) was added over 20 minutes to a stirredsolution oftert-butyl((2R)-1-(1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)propan-2-yl)carbamate(1.27 Kg, 1.76 mol) in IPA (1.50 L), keeping the internal temperaturebelow 30° C. The resulting mixture was heated to 40° C. and stirred at40° C. for 16 hours. The temperature was increased to 55° C. and afurther portion of 12 M HCl (0.20 L, 2.40 mol) was added and stirringwas continued for 7 hours. MTBE (2.50 L) was added and the mixture wasallowed to stir at 25° C. for 16 hours. The resulting solid wascollected by filtration and the filter-cake was washed with 10% IPA inMTBE (2.00 L). The resulting solid was dried under reduced pressure at40° C. for 72 hours to give(2R)-1-(1H-indazol-4-yl)propan-2-amine;dihydrochloride (0.39 Kg, 89%) asan off-white solid.

¹H NMR (500 MHz, DMSO, 27° C.) 1.12 (3H, d), 2.96 (1H, dd), 3.39 (1H,dd), 3.45-3.55 (1H, m), 6.94 (1H, d), 7.28 (1H, dd), 7.43 (1H, d), 8.23(3H, s), 8.28 (1H, d). m/z: ES+ [M.+] 175.

Preparation of(2R)-1-(1H-indazol-4-yl)-N-(2,2,2-trifluoroethyl)propan-2-amine

A solution of 2,2,2-trifluoroethyl trifluoromethanesulfonate (0.73 Kg,2.94 mol) in MeCN (2.00 L) was added to a stirred mixture of(R)-1-(1H-indazol-4-yl)propan-2-amine dihydrochloride (0.77 Kg, 2.94mol) and K₂CO₃ (1.26 Kg, 9.13 mol) in MeCN (8.00 L) at 25° C. undernitrogen. The resulting mixture was heated to 60° C. and stirred at 60°C. for 18 hours. MeCN (5.00 L) was added and the solids were removed byfiltration. The filtrate was concentrated under reduced pressure and theresulting solid was dissolved in EtOAc (10.00 L), washed with water (4.0L) and dilute saturated aqueous sodium chloride (4.8 L). The organiclayer was concentrated under reduced pressure to give(2R)-1-(1H-indazol-4-yl)-N-(2,2,2-trifluoroethyl)propan-2-amine (0.73Kg, 96%). ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.12 (3H, d), 2.96 (1H, dd),3.08 (1H, dd), 3.14-3.33 (3H, m), 6.98 (1H, d), 7.32 (1H, dd), 7.37-7.41(1H, m), 8.13 (1H, d), 10.86 (1H, s). m/z: ES+ [M+H]+ 257.

Preparation of(6S,8R)-6-(5-bromo-2-pyridyl)-8-methyl-7-(2,2,2-trifluoroethyl)-3,6,8,9-tetrahydropyrazolo[4,3-f]isoquinoline

Trifluoroacetic acid (0.66 L, 8.66 mol) was slowly added to a stirredmixture of 5-bromopicolinaldehyde (0.55 Kg, 2.89 mol) and(R)-1-(1H-indazol-4-yl)-N-(2,2,2-trifluoroethyl)propan-2-amine (0.83 Kg,2.89 mol) in toluene (7.30 L) at 21° C. The resulting solution washeated to 80° C. and stirred at 80° C. for 18 hours. The temperature wasincreased to 90° C. and heating continued for 4 hours. The mixture wasallowed to cool to 80° C. and a further portion of5-bromopicolinaldehyde (0.015 Kg, 0.081 mol) was added and the solutionwas stirred at 80° C. for 16 hours. The mixture was allowed to cool to21° C. and saturated aqueous NaHCO₃ (6.60 L) was added over 20 minutes.EtOAc (5.00 L) was added and the layers were separated. The organiclayer was washed with saturated aqueous NaHCO₃ (3.30 L) and concentratedunder reduced pressure. Ethanol was added and the mixture wasconcentrated under reduced pressure to give the crude product as acrispy solid. The cis isomer was removed by preparative SFC (SuperSep 1™column (filled with CelluCoat™ 10 μm), 50 mm diameter, 250 mm length),eluting 28% EtOH in CO₂, 140 bar mobile phase at a flow rate of 450mL/minute at 30° C.) to give to give(6S,8R)-6-(5-bromo-2-pyridyl)-8-methyl-7-(2,2,2-trifluoroethyl)-3,6,8,9-tetrahydropyrazolo[4,3-f]isoquinoline(0.94 Kg, 77%) as a pale yellow solid. ¹H NMR (500 MHz, CDCl₃, 27° C.)1.16 (3H, d), 2.89 (1H, dd), 2.99 (1H, dq), 3.25-3.34 (2H, m), 3.55 (1H,td), 5.10 (1H, s), 6.93 (1H, d), 7.22 (1H, d), 7.40 (1H, d), 7.75 (1H,dd), 8.05 (1H, d), 8.56 (1H, dd), 10.24 (1H, s). m/z: ES+ [M+H]+ 427.

Alternative Route #2 to Example 17 Preparation ofN-[1-(3-fluoropropyl)azetidin-3-yl]-6-[(6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-3,6,8,9-tetrahydropyrazolo[4,3-f]isoquinolin-6-yl]pyridin-3-amine

1-(3-Fluoropropyl)azetidin-3-amine (6.31 g, 37.98 mmol) was added to asolution of (6S,8R)-6-(5-bromopyridin-2-yl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline (15.47 g,34.92 mmol) in 1,4-dioxane (175 mL). The resulting solution was degassedunder vacuum for 5 minutes, then backfilled with nitrogen (×2).

Sodium tert-butoxide (13.43 g, 139.69 mmol) was added, followed byBrettPhos 3rd Generation Precatalyst (0.95 g, 1.05 mmol). The resultingmixture was heated at 55° C. for 18 hours. The mixture was poured intoEtOAc and water. Saturated brine was added, and the layers wereseparated. The organic layer was washed with saturated aqueous sodiumchloride (×3), and the combined aqueous layers were extracted withEtOAc. The combined organic layers were dried over Na₂SO₄, filtered andconcentrated under reduced pressure. The resulting residue was purifiedby flash silica chromatography, elution gradient 0 to 50% methanol inethyl acetate. Fractions containing the desired product were combined,concentrated under reduced pressure, and then repurified by flash silicachromatography, elution gradient 0 to 40% EtOAc in hexanes, to afford ared-orange solid. This solid was dissolved in 10% MeOH in DCM, filtered,washing the filtercake with DCM. The combined filtrates wereconcentrated under reduced pressure to giveN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine(15.28 g, 92%) as a light orange foam solid. The trans and cis isomerswere separated by preparative SFC ((S,S) Whelk-01 column, 30 mmdiameter, 250 mm length), eluting with 30% (0.2% NH₄OH in MeOH) in CO₂,100 bar mobile phase at a flow rate of 20 mL/minute at 40° C.). Thefractions containing the trans isomer were concentrated under reducedpressure. The resulting residue was further purified by flash silicachromatography, elution gradient 0 to 30% MeOH in EtOAc. Fractionscontaining the desired product were concentrated under reduced pressure,then concentrated from MeCN (×2) to giveN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine(10.76 g, 74%) as a pale yellow foam/solid. ¹H NMR (600 MHz, DMSO-d₆,27° C.) 1.07 (3H d) 1.63 (2H, dquin), 2.45 (2H, t), 2.72 (2H, br t),2.83 (1H, br dd), 2.91-3.00 (1H, m), 3.06 (1H, br dd) 3.42-3.55 (2H, m),3.58-3.65 (2H, m), 3.92 (1H, dquin) 4.43 (2H, dt) 4.92 (1H, s), 6.22(1H, d), 6.79 (1H, d) 6.82 (1H, dd), 6.96 (1H, d), 7.21 (1H, d), 7.73(1H, d), 8.04 (1H, s), 12.96 (1H, s). m/z: ES+(M+H)+477.

Fractions from the SFC containing the cis isomer was concentrated underreduced pressure and the resulting residue was further purified by flashsilica chromatography, elution gradient 0 to 30% MeOH in EtOAc.Fractions containing the desired product were concentrated under reducedpressure, and the resulting residue was filtered to affordN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6R,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine(1.13 g, 8%) as a light yellow foam solid. ¹H NMR (300 MHz, DMSO-d₆, 27°C.) 1.28 (3H, d), 1.55-1.75 (2H, m), 2.42-2.49 (2H, m), 2.76 (2H, br t),2.80-2.92 (1H, m), 3.07-3.18 (1H, m), 3.18-3.28 (1H, m), 3.38-3.54 (2H,m), 3.59-3.70 (2H, m), 3.95 (1H, sxt), 4.45 (2H, dt), 5.11 (1H, s), 6.21(1H, d), 6.72 (1H, d), 6.82 (1H, dd), 7.02 (1H, d), 7.19 (1H, d), 7.78(1H, d), 8.06 (1H, s), 12.94 (1H, s). m/z: ES+(M+H)+477.

Procedures used to prepare the starting material(6S,8R)-6-(5-bromopyridin-2-yl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineare described below.

Preparation of2,2,2-trifluoro-N-[(1R)-2-(1H-indazol-4-yl)-1-methyl-ethyl]acetamide

TEA (26.0 mL, 186.5 mmol) was added via syringe to a stirred slurry of(R)-1-(1H-indazol-4-yl)propan-2-amine dihydrochloride (15.29 g, 61.61mmol) and ethyl 2,2,2-trifluoroacetate (8.09 mL, 67.8 mmol) in MeOH (150mL) at room temperature. The resulting solution was stirred at roomtemperature for 18 hours then concentrated under reduced pressure. Theresulting solid was then dissolved in DCM (40 mL) and the residualtriethylamine hydrochloride was precipitated out by the addition Et₂O(300 mL). The slurry was stirred for 5 minutes then filtered and thefiltrate was concentrated under reduced pressure to give a solid. Thesolid was dissolved in EtOAc and saturated aqueous sodium chloride. Thelayers were separated and the organic layer was washed with saturatedsaturated aqueous sodium chloride, dried over MgSO₄, filtered andconcentrated to afford the(R)—N-(1-(1H-indazol-4-yl)propan-2-yl)-2,2,2-trifluoroacetamide (16.51g, 99%) as an off white solid. ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 1.19(3H, d), 3.03 (1H, dd), 3.13 (1H, dd), 4.13-4.30 (1H, m), 6.89 (1H, d),7.25 (1H, dd), 7.38 (1H, d), 8.18 (1H, t), 9.35 (1H, br d), 13.01 (1H,s). m/z: ES+[M+H]+ 272.

Preparation of(2R)-1-(1H-indazol-4-yl)-N-(2,2,2-trifluoroethyl)propan-2-amine

1M Borane THF complex in THF (365 mL, 365.0 mmol) was added rapidlydropwise via cannula to a stirred solution of(R)—N-(1-(1H-indazol-4-yl)propan-2-yl)-2,2,2-trifluoroacetamide (16.5 g,60.8 mmol) in THF (100 mL) at room temperature (gas evolution; smallexotherm observed during course of addition). The reaction was thenheated at 60° C. for 15.5 hours then allowed to cool to roomtemperature. The reaction was placed in an ambient temperature waterbath and MeOH (80 mL) was slowly added drop-wise. The reaction mixturewas concentrated under reduced pressure, and the resulting residue wasdissolved in MeOH (80 mL) and concentrated under reduced pressure. Theresulting residue was dissolved in MeOH (250 mL) and placed in anambient temperature water bath. A slurry of 10% palladium on carbon(6.47 g, 6.08 mmol) in MeOH (90 mL) was added, rinsing down the flaskwith MeOH (10 mL) (some gas evolution was noted). The mixture wasstirred at room temperature for approximately 3 minutes then heated at65° C. for 2 hours, After cooling to room temperature, the mixture wasfiltered through celite and concentrated under reduced pressure. Thepale yellow residue was dissolved in DCM and filtered and the filtratewas concentrated under reduced pressure. The resulting residue waspurified by flash silica chromatography, elution gradient 10 to 60%EtOAc in hexanes. Fractions containing the desired product wereconcentrated under reduced pressure to afford(R)-1-(1H-indazol-4-yl)-N-(2,2,2-trifluoroethyl)propan-2-amine (14.21 g,86%) as a pale yellow oil. ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 0.91 (3H,d), 2.17-2.30 (1H, m), 2.71 (1H, dd), 2.98-3.17 (2H, m), 3.22-3.38 (2H,m), 6.89 (1H, d), 7.24 (1H, dd), 7.35 (1H, d), 8.12 (1H, t), 12.97 (1H,br s). m/z: ES+ [M+H]+ 258.

Preparation of(6S,8R)-6-(5-bromo-2-pyridyl)-8-methyl-7-(2,2,2-trifluoroethyl)-3,6,8,9-tetrahydropyrazolo[4,3-f]isoquinoline

5-Bromopicolinaldehyde (9.80 g, 52.7 mmol) was added to a stirredsolution of(R)-1-(1H-indazol-4-yl)-N-(2,2,2-trifluoroethyl)propan-2-amine (14.05 g,51.66 mmol) in toluene (246 mL) under nitrogen. Trifluoroacetic acid(12.30 mL) was then added and the reaction was heated at 90° C. for 4.5hours. The mixture was allowed to cool to room temperature and EtOAc(200 mL) was added. The mixture was cooled to 5° C. and the mixture wasbasified by slow addition of saturated aqueous NaHCO₃. The resultinglayers were separated and the aqueous layer was extracted with EtOAc(2×80 mL). The combined organic extracts were washed with saturatedaqueous NaHCO₃, saturated aqueous sodium chloride, dried over MgSO₄,filtered and concentrated under reduced pressure. The resulting residuewas purified by flash silica chromatography, elution gradient 5 to 60%EtOAc in hexanes. Fractions containing the desired product wereconcentrated under reduced pressure, then dried in vacuo at 50° C. for 3hours to afford(6S,8R)-6-(5-bromopyridin-2-yl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(19.29 g, 88%) as a pale orange solid. 1H NMR analysis revealed a 10:1trans/cis isomers. ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 1.10 (3H, d),2.83-2.94 (1H, m), 2.95-3.14 (2H, m), 3.34-3.47 (1H, m), 3.49-3.72 (1H,m), 5.10 (1H, s), 6.88 (1H, d), 7.25 (1H, d), 7.33 (1H, d), 7.99 (1H,dd), 8.06 (1H, d), 8.56 (1H, d), 13.00 (1H, s) m/z: ES+ [M+H]+ 425.

Alternative routes to intermediates used in the synthesis of Example 17:

Preparation ofN-[(1R)-2-(3-amino-2-methyl-phenyl)-1-methyl-ethyl]-2,2,2-trifluoro-acetamide

A solution of (R)-3-(2-aminopropyl)-2-methylaniline (0.51 g, 3.13 mmol),DIPEA (0.55 mL, 3.13 mmol), and ethyl 2,2,2-trifluoroacetate (0.47 mL,3.91 mmol) in MeOH (9.5 mL) was stirred at room temperature undernitrogen for 16 hours. The reaction mixture was concentrated underreduced pressure. DCM and water were added and the layers wereseparated. The aqueous layer was extracted with DCM (3×50 mL), and thecombined organic layers were dried (Na₂SO₄) and filtered. The crudeproduct was purified by flash silica chromatography, elution gradient 0to 100% EtOAc in hexane. Pure fractions were evaporated to dryness toaffordN-[(1R)-2-(3-amino-2-methyl-phenyl)-1-methyl-ethyl]-2,2,2-trifluoro-acetamide(680 mg, 83%). ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 1.06-1.14 (m, 3H), 2.01(s, 3H), 2.54-2.71 (m, 1H), 2.72-2.84 (m, 1H), 4.04-4.12 (m, 1H),4.60-4.77 (m, 2H), 6.24-6.37 (m, 1H), 6.43-6.52 (m, 1H), 6.72-6.84 (m,1H), 9.19-9.32 (m, 1H). m/z: ES+ [M+H]+ 261.

Preparation of2-methyl-3-[(2R)-2-(2,2,2-trifluoroethylamino)propyl]aniline

1.0M borane in THF, in THF (19.4 mL, 19.37 mmol) was added to a stirredsolution of(R)—N-(1-(3-amino-2-methylphenyl)propan-2-yl)-2,2,2-trifluoroacetamide(840 mg, 3.23 mmol) in THF (10 mL). The reaction mixture was heated at65° C. under nitrogen for 6 hours. The mixture was allowed to cool toroom temperature and MeOH was carefully added. The resulting mixture wasstirred at room temperature for 30 minutes and then concentrated underreduced pressure. The resulting residue was dissolved in MeOH and heatedto 65° C. for 6 hours. The mixture was allowed to cool to roomtemperature and then concentrated under reduced pressure to give thecrude product which was purified by flash silica chromatography, elutiongradient 0 to 100% EtOAc in hexane. Product containing fractions wereevaporated to dryness to afford2-methyl-3-[(2R)-2-(2,2,2-trifluoroethylamino)propyl]aniline (665 mg,84%). ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 0.89 (s, 3H), 1.98 (s, 3H),2.08-2.22 (m, 1H), 2.25-2.42 (m, 1H), 2.69-2.87 (m, 2H), 3.15-3.29 (m,2H), 4.61-4.75 (m, 2H), 6.30-6.40 (m, 1H), 6.43-6.54 (m, 1H), 6.71-6.83(m, 1H). m/z: ES+ [M+H]+ 247.

Preparation of(2R)-1-(3-bromo-2-methyl-phenyl)-N-(2,2,2-trifluoroethyl)propan-2-amine

DIPEA (2.99 mL, 17.10 mmol) was added to(R)-1-(3-bromo-2-methylphenyl)propan-2-amine (1.30 g, 5.70 mmol), and2,2,2-trifluoroethyl trifluoromethanesulfonate (0.23 M in DCM, 29.7 mL,6.84 mmol) in 1,4-dioxane (25 mL) at room temperature under nitrogen.The resulting mixture was stirred at 85° C. for 16 hours. The reactionmixture was concentrated under reduced pressure and the crude productwas purified by flash silica chromatography, elution gradient 0 to 50%EtOAc in heptane. Product containing fractions were concentrated underreduced pressure to afford(R)-1-(3-bromo-2-methylphenyl)-N-(2,2,2-trifluoroethyl)propan-2-amine(0.56 g, 32%) as an oil. ¹H NMR (500 MHz, DMSO, 27° C.) 0.91 (3H, d),2.26 (1H, q), 2.34 (3H, s), 2.78-2.86 (1H, m), 2.89 (1H, dd), 3.19-3.28(2H, m), 7.01-7.06 (1H, m), 7.15 (1H, dd), 7.43 (1H, dd). m/z: ES+[M+H]+ 310/312.

Preparation of2-methyl-3-[(2R)-2-(2,2,2-trifluoroethylamino)propyl]aniline

Tris(dibenzylideneacetone)dipalladium(0) (0.050 g, 0.05 mmol) and(±)-2,2′-Bis(diphenylphosphino)-1,1′-binaphthalene (0.067 g, 0.11 mmol)were added to a suspension of(R)-1-(3-bromo-2-methylphenyl)-N-(2,2,2-trifluoroethyl)propan-2-amine(0.56 g, 1.81 mmol), benzophenone imine (0.33 mL, 1.99 mmol) and sodiumtert-butoxide (0.26 g, 2.71 mmol) in degassed toluene (7 mL) and thereaction was heated to 90° C. for 16 hours. After cooling to roomtemperature, the mixture was concentrated under reduced pressure. Theresidue was dissolved in DCM (50 mL) and washed with water (50 mL). Theaqueous was extracted with DCM (25 mL) and the combined organics wereconcentrated to ˜25 mL. 2 M HCl solution (50 mL) was added and thebiphasic mixture was stirred vigorously for 30 min. The layers wereseparated and the aqueous layer was washed with DCM. The aqueous phasewas basified by addition of 2 M aqueous NaOH. The resulting mixture wasextracted with DCM (2×100 mL) and the combined organic extracts wereconcentrated under reduced pressure to give(R)-2-methyl-3-(2-((2,2,2-trifluoroethyl)amino)propyl)aniline (0.31 g,70%) as an oil. ¹H NMR (500 MHz, DMSO, 27° C.) 0.90 (3H, d), 1.98 (3H,s), 2.12-2.18 (1H, m), 2.30-2.36 (1H, m), 2.74-2.82 (2H, m), 3.17-3.29(2H, m), 4.69 (2H, s), 6.35 (1H, dd), 6.48 (1H, dd), 6.78 (1H, t). ES+[M+H]+247.

Preparation of(1S,3R)-1-(5-bromo-2-pyridyl)-3,5-dimethyl-2-(2,2,2-trifluoroethyl)-3,4-dihydro-1H-isoquinolin-6-amine

Trifluoromethanesulfonic acid ytterbium(III) salt (0.17 g, 0.27 mmol)was added to a stirred solution of(R)-2-methyl-3-(2-((2,2,2-trifluoroethyl)amino)propyl)aniline (1.35 g,5.49 mmol), 5-bromopicolinaldehyde (1.02 g, 5.49 mmol) and water (0.49mL, 27.43 mmol) in MeCN (21.5 mL). The resulting solution was stirred at80° C. for 1 hour. The mixture was concentrated under reduced pressureand the residue was dry-loaded onto silica gel and was purified by flashsilica chromatography, elution gradient 10 to 30% EtOAc in hexane.Fractions containing the desired product were concentrated to dryness toafford(1S,3R)-1-(5-bromopyridin-2-yl)-3,5-dimethyl-2-(2,2,2-trifluoroethyl)-1,2,3,4-tetrahydroisoquinolin-6-amine(2.22 g, 98%) as a yellowish gum. ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 0.88(3H, d), 1.91 (3H, s), 2.37-2.46 (1H, m), 2.60-2.76 (1H, m), 2.76-3.00(1H, m), 3.15-3.28 (1H, m), 3.34-3.63 (1H, m), 4.65 (2H, s), 4.85 (1H,s), 6.19-6.52 (2H, m), 7.10-7.31 (1H, m), 7.97 (1H, dd), 8.62 (1H, d).m/z: ES+ [M+H]+ 414.

Preparation of(6S,8R)-6-(5-bromo-2-pyridyl)-8-methyl-7-(2,2,2-trifluoroethyl)-3,6,8,9-tetrahydropyrazolo[4,3-f]isoquinoline

(1S,3R)-1-(5-bromopyridin-2-yl)-3,5-dimethyl-2-(2,2,2-trifluoroethyl)-1,2,3,4-tetrahydroisoquinolin-6-amine(2.21 g, 5.33 mmol) in propionic acid (14.0 mL) was cooled to −20° C. Asolution of sodium nitrite (0.40 g, 5.86 mmol) in water (2.8 mL) wasadded dropwise over 5 minutes, keeping the bath temperature between −20°C. and −15° C. The resulting mixture was stirred at −20° C. for 45minutes. Toluene (80 mL), cooled to −15° C., was poured into thereaction, and the reaction was stirred for 15 minutes. A solution ofsodium carbonate (12.0 g, 113 mmol) in water (125 mL) was slowly addedand the mixture was allowed to warm to room temperature. EtOAc (100 mL)was added and the layers were separated. The aqueous layer was extractedwith CHCl₃/IPA (3:1) (100 mL). The combined organic layers were driedover MgSO₄, filtered, and concentrated under reduced pressure to givethe crude product which was purified by flash silica chromatography,elution gradient 15 to 50% EtOAc in hexane. Product containing fractionswere evaporated to dryness to afford(6S,8R)-6-(5-bromopyridin-2-yl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(1.63 g, 72%) as a brown solid. ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 1.10(3H, d), 2.83-2.96 (1H, m), 2.96-3.14 (2H, m), 3.35-3.48 (1H, m),3.49-3.73 (1H, m), 5.11 (1H, s), 6.87 (1H, d), 7.27 (1H, d), 7.35 (1H,d), 7.99 (1H, dd), 8.09 (1H, s), 8.53-8.63 (1H, m), 12.96 (1H, s). m/z:ES+ [M+H]+ 425.

Preparation of tert-butyl (1-Q-fluoropropyl)azetidin-3-yl)carbamate

A suspension of tert-butyl azetidin-3-ylcarbamate (10.00 g, 58.06 mmol),1-fluoro-3-iodopropane (11.13 g, 59.22 mmol) and potassium carbonate(16.05 g, 116.13 mmol) in MeCN (200 mL) were stirred at room temperaturefor 2 days. The solids were removed by filtration and the filtrate wasconcentrated under reduced pressure. The residue was treated with waterand extracted with DCM. The combined organic extracts were dried overNa₂SO₄, filtered and concentrated under reduced pressure to givetert-butyl (1-(3-fluoropropyl)azetidin-3-yl)carbamate (13.40 g, 99%) asa pale yellow solid. ¹H NMR (300 MHz, CDCl₃, 27° C.) 1.45 (9H, s),1.61-1.92 (2H, m), 2.51-2.67 (2H, m), 2.85 (2H, br s), 3.57-3.72 (2H,m), 4.30 (1H, br d), 4.36-4.63 (2H, m), 4.89 (1H, br d).

Alternative Preparation of tert-butyl(1-(3-fluoropropyl)azetidin-3-yl)carbamate

Trifluoromethanesulfonic anhydride (2.31 mL, 13.75 mmol), followed by2,6-dimethylpyridine (1.74 mL, 15.00 mmol) were added to a solution of3-fluoropropan-1-ol (0.94 mL, 12.5 mmol) in DCM (47 mL) at 0° C. and thereaction was stirred at 0° C. for 1 hour. The reaction mixture waswashed with 1N HCl, then the organic layer was dried and carefullyevaporated. The residue was then added to a separate flask containing asuspension of tert-butyl azetidin-3-ylcarbamate (1.94 g, 11.25 mmol) andDIPEA (3.24 ml, 18.75 mmol) in dioxane/DCM (1:1, 40 mL) and the reactionwas stirred at room temperature (exothermic on addition of triflate).The reaction mixture was diluted with DCM and washed with NH₄Clsolution. The organic layer was dried and evaporated under reducedpressure to give the crude product. The crude product was purified byflash silica chromatography, elution gradient 0 to 10% 1M NH₃/MeOH inDCM. Pure fractions were evaporated to dryness to afford tert-butyl(1-(3-fluoropropyl)azetidin-3-yl)carbamate (2.82 g, 97%) as a mauve oil¹H NMR as above.

Preparation of 1-(3-fluoropropyl)azetidin-3-amine

Trifluroacetic acid (39.8 mL, 517 mmol) was added dropwise over 30minutes to a solution of tert-butyl(1-(3-fluoropropyl)azetidin-3-yl)carbamate (20.0 g, 86 mmol) in DCM(67.8 mL) that had been immersed in a water bath. After 18 hours, thereaction was concentrated under reduced pressure to an orange oil. Thisoil was dissolved in 10% methanol in DCM (125 mL) and potassiumcarbonate (71.4 g, 517 mmol) was added with vigorous stirring. After 15min, another 30 g of potassium carbonate were added. Stirring becamesluggish, and Celite was added (˜15 g). Stirring was continued foranother 15 minutes, and the mixture was filtered with a 10% methanol inDCM wash. The filtrate was concentrated under reduced pressure on arotary evaporator (pressure: 100 mbar, water bath temperature: 30° C.),and the resulting orange oil was purified by flash silica chromatography(λ detection=210 nm), elution gradient 0 to 15% (2 M ammonia inmethanol) in DCM to afford 1-(3-fluoropropyl)azetidin-3-amine (12.8 g)contaminated with 28 wt % methanol based on NMR integration as a yellowoil. This material could be used directly, stored in the freezer forlater use, or further purified by vacuum distillation as according tothe following example: 1-(3-fluoropropyl)azetidin-3-amine (14.8 g) as alight yellow oil was distilled using a short path distillation apparatuswith jacketed water cooling under vacuum (vacuum pump) conditions. Theflask containing the amine was immersed in an oil bath, and the bathtemperature was gradually increased to 140° C. over a period of 30minutes. Distillation occurred with a bath temperature of 110-135° C.and an approximate head temperature of 70° C. to afford1-(3-fluoropropyl)azetidin-3-amine (12.4 g, 84%) as a clear colorlessoil. This oil was stored in the freezer before use. ¹H NMR (300 MHz,DMSO-d₆, 27° C.) 1.52-1.70 (2H, m), 1.75 (2H, br s), 2.38 (2H, t),2.43-2.48 (2H, m), 3.27-3.39 (1H, m), 3.42-3.48 (2H, m), 4.43 (2H, dt).m/z: ES+ [M+H]+ 133.

Example 17A: Preparation ofN-(1-(3-fluoropropyl)azetidin-3-yl)-64(6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amineForm A (anhydrous form) Method 1

4.0 mg of amorphousN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-aminewas dissolved in 100 ul of EtOAc to form a clear solution. About 200 ulof heptane was added slowly to the solution, a cloudy suspension wasformed. The slurry was stirred at the ambient condition for 3 days.Crystalline material of Form A was obtained.

Method 2

1.60 g of amorphousN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-aminewas suspended in 15 ml of acetone/H₂O (10:1) mixture. A slurry wasformed, and then a gel was formed. 10 mg ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amineForm A seed was added after 2 hours, and the suspension was stirred atthe ambient temperature for 1 day whereupon the gel became solid cakes.The solid cakes on the wall and bottom were manually removed into thesolution, and a homogenous slurry was obtained after stirring for 1hour. The solid was collected by filtration and washed with 5.0 ml ofacetone/H₂O (1:10) twice, followed with H₂O several times. 1.45 g ofwhite powder was obtained (90% yield) as Form A after drying in vacuumfor 4 hours.

Method 3

501 mg of amorphous material ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-aminewas dissolved in 1.0 ml of EtOAc to form a brown solution. To thesolution, 1.0 ml of heptane was added slowly. At the end of theaddition, solid started to form but then dissolved. 2-5 mg ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine-FormA was added to the clear brown solution, whereupon solid started toprecipitate. The suspension was stirred at the ambient temperature for 1day. The solid was collected by filtration and dried in air. 345 mg ofoff-white powder was obtained (yield: ˜69%) as Form A.

Form A from method 2 was analyzed by XRPD and the results are tabulatedbelow (Table 1) and shown in FIG. 1.

TABLE 1 XRPD Peaks for Form A Angle (2θ ± 0.2°) Intensity (%) 21.1 100.020.8 54.3 14.6 41.9 18.6 41.6 12.3 38.9 15.5 34.1 24.6 31.3 15.8 30.613.4 23.2 19.0 21.7

Form A was analyzed by thermal techniques. DSC analysis indicated thatForm A has a melting point with an onset at 132° C. and a peak at 137°C. TGA indicated that Form A exhibits a mass loss of about 0.2% uponheating from about 25° C. to about 100° C. A representative DSC/TGAthermogram of Form A is shown in FIG. 2.

Example 17B: Preparation ofN-(1-(3-fluoropropyl)azetidin-3-yl)-64(6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amineForm B (TBME solvate)

100 mg ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine(amorphous material) was partially dissolved in 1.0 ml of TBME. Thesuspension was stirred under ambient conditions, whereupon more solidprecipitated out in the suspension. The slurry was stirred for 2 hours.N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine-FormB was identified when the slurry was directly amounted in the sampleholder for the XRPD characterization. Form B converted to Form C afterthe solid was filtered and dried in the ambient condition.

Form B was analyzed by XRPD and the results are tabulated below (Table2) and shown in FIG. 3.

TABLE 2 XRPD Peaks for Form B Angle (2θ ± 0.2°) Intensity (%) 6.0 100.017.8 67.0 18.4 34.9 16.3 30.3 21.5 29.2 12.3 27.0 17.2 24.8 13.4 23.318.1 21.6 15.6 20.6

Example 17C: Preparation ofN-(1-(3-fluoropropyl)azetidin-3-yl)-64(6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amineForm C (TBME solvate)

200 mg of N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amineamorphous material was almost dissolved in 1 ml of TBME. The suspensionwas stirred under ambient conditions, whereupon more solid precipitatedout after 10 minutes. The slurry was stirred for 2 hours and thenevaporated under ambient conditions. Form C was obtained after theresulting solid was dried under ambient conditions.

Form C was analyzed by XRPD and the results are tabulated below (Table3) and shown in FIG. 4.

TABLE 3 XRPD Peaks for Form C Angle (2θ ± 0.2°) Intensity (%) 6.8 100.018.6 64.1 13.9 62.5 18.0 61.6 17.2 53.4 17.4 52.4 20.5 47.3 19.2 40.020.7 37.4 13.7 36.3

Form C was analyzed by thermal techniques. DSC analysis indicated thatForm C has an endotherm event of desolvation with an onset at 75° C. anda peak at 84° C. TGA indicated that Form C exhibits a mass loss of about7.1% upon heating from about 25° C. to about 150° C. A representativeDSC/TGA thermogram of Form C is shown in FIG. 5.

Example 17D: Preparation ofN-(1-(3-fluoropropyl)azetidin-3-yl)-64(6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amineForm D (CPME solvate)

150 mg of N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amineamorphous material was stirred in 2.0 ml of CPME (cyclopentyl methylether). After stirring for 1 day, the solid was isolated by filtrationand dried in air. ˜50 mg of white powder of Form D was obtained.

Form D was analyzed by XRPD and the results are tabulated below (Table4) and shown in FIG. 6.

TABLE 4 XRPD Peaks for Form D Angle (2θ ± 0.2°) Intensity (%) 6.2 100.018.3 55.0 18.9 24.3 16.1 21.8 16.8 18.4 22.2 17.9 14.1 17.1 19.9 13.522.5 13.5 23.9 13.4

Form D was analyzed by thermal techniques. DSC analysis indicated thatForm D has an endotherm event of desolvation with an onset at 76° C. anda peak at 81° C., followed by another endotherm event with an onset at128° C. and a peak at 133° C. TGA indicated that Form D exhibits a massloss of about 6.8% upon heating from about 25° C. to about 150° C. Arepresentative DSC/TGA thermogram of Form D is shown in FIG. 7.

Example 17E: Preparation ofN-(1-(3-fluoropropyl)azetidin-3-yl)-64(6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amineForm E (anhydrous form) Method 1

100 mg of amorphous N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-aminewas added into 2.0 ml of heptane to form a slurry. 0.40 ml of EtOAc wasadded. After stirring for 2 hours, extra 0.10 ml of EtOAc was added tothe slurry and the slurry was stirred under ambient conditions for 18hours, whereupon a new crystalline form, Form E, was identified by XRPD.75 mg of white solid of Form E was obtained after the solid was isolatedand dried at the ambient condition (yield: 75%).

Method 2

1.002 g of amorphousN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-aminewas dissolved in 5.0 ml of EtOAc to obtain a clear light brown solution.5.0 ml of heptane was added and the solution remained clear. 10 mg ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine-FormE seeds were added, and the suspension was stirred at ambienttemperature. The solid started to precipitate under ambient conditions,and a wet cake was formed after 1 hour. 5.0 ml of EtOAc was added slowlyto the slurry and the resulting slurry was stirred at ambienttemperature for 1 hour. Another 10 ml of heptane was added, more solidstarted to precipitate. After stirring under ambient conditions for 2hours, the slurry was heated to 60° C. and stirred at 60° C. for 2hours. The slurry was cooled down to ambient temperature and stirred for18 hours. The off-white solid of Form E was isolated by filtration andwashed with 1:4 EtOAc/heptane mixed solvent for 3 times. 0.875 g of palewhite powder was collected after air-dried (yield: 87%).

Form E from method 1 was analyzed by XRPD and the results are tabulatedbelow (Table 5) and shown in FIG. 8.

TABLE 5 XRPD Peaks for Form E Angle (2θ ± 0.2°) Intensity (%) 17.9 100.014.8 67.1 20.9 60.1 23.1 55.4 16.2 49.3 20.0 35.6 18.2 32.9 12.3 30.422.2 19.0 13.9 18.9

Form E was analyzed by thermal techniques. DSC analysis indicated thatForm E has a melting point with an onset at 126° C. and a peak at 133°C. TGA indicated that Form E exhibits a mass loss of about 0.8% uponheating from about 25° C. to about 100° C. A representative DSC/TGAthermogram of Form E is shown in FIG. 9.

Example 17F: Preparation ofN-(1-(3-fluoropropyl)azetidin-3-yl)-64(6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amineForm F (heptane solvate)

20 mg of amorphousN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-aminewas dissolved in 200 ul of acetone. 1.0 ml of heptane was added slowlyto the solution. The solution started to become cloudy and then formed aslurry. The slurry was stirred at the ambient temperature after 1.0 mlof heptane was added. Needle-like particles was observed in the slurry.The slurry was heated to 60° C. and stirred for 2 hours and then cooleddown to the ambient temperature. Needle particles were formed.

The slurry of resulting Form F was analyzed by XRPD and the results areshown in FIG. 10. Crystallinity of Form F was decreasing during XRPDcharacterization, and the second run of the XRPD indicated an amorphousmaterial was obtained after drying in the sample holder.

Example 17F: Preparation ofN-(1-(3-fluoropropyl)azetidin-3-yl)-64(6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amineForm G (methylpentanone solvate)

100 mg of amorphous N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-aminewas suspended in 200 ul of methylpentanone, and 2-5 mg ofN-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine-FormA was added, whereupon a cake was formed after stirring under ambientconditions for 10 minutes. 200 ul of methylpentanone was added and aslurry was formed. The slurry was stirred at the ambient condition for18 hours. XRPD showed that a crystalline form,N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine-FormG was formed. Form G converted to partially amorphous material afterisolation and drying under ambient conditions.

Form G was analyzed by XRPD and the results are tabulated below (Table6) and shown in FIG. 10.

TABLE 6 XRPD Peaks for Form G Angle (2θ ± 0.2°) Intensity (%) 6.2 100.018.1 95.9 16.6 38.8 18.7 37.5 21.9 36.5 15.9 33.9 22.4 28.1 19.7 26.414.0 24.6 12.6 23.9

Example 185-fluoro-6-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine

[(2-Di-cyclohexylphosphino-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl)-2-(2′-amino-1,1′-biphenyl)]palladium(II)methanesulfonate methanesulfonate (Brett Phos G3) (48.6 mg, 0.05 mmol)was added to degassed solution of 1-(3-fluoropropyl)azetidin-3-amine (95mg, 0.72 mmol),(6S,8R)-6-(5-bromo-3-fluoropyridin-2-yl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(155 mg, 0.36 mmol) and sodium tert-butoxide (206 mg, 2.15 mmol) in1,4-dioxane (3 mL) under nitrogen. The resulting solution was stirred at90° C. for 2 hours. The reaction mixture was diluted with EtOAc (50 mL),and washed sequentially with water (50 mL) and saturated aqueous sodiumchloride (50 mL). The organic layer was dried with MgSO₄, filtered andevaporated to afford crude product which was purified by preparativeHPLC (Waters XBridge Prep C18 OBD column, 5μ silica, 50 mm diameter, 100mm length), using decreasingly polar mixtures of water (containing 0.1%NH₃) and MeCN as eluents. Fractions containing the desired compound wereevaporated to dryness to afford5-fluoro-6-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine(39.0 mg, 22%) as a white solid. ¹H NMR (500 MHz, CDCl₃, 27° C.)0.48-0.61 (2H, m), 0.92-1.09 (2H, m), 1.13 (3H, d), 1.68-1.83 (2H, m),2.56-2.62 (2H, m), 2.67 (1H, dd), 2.86 (1H, dd), 2.90-2.95 (2H, m), 3.17(1H, dd), 3.27 (1H, dd), 3.70 (2H, q), 3.87 (1H, td), 4.04 (1H, q), 4.21(1H, d), 4.43 (1H, t), 4.53 (1H, t), 5.39 (1H, s), 6.50 (1H, dd), 6.79(1H, d), 7.12 (1H, d), 7.68 (1H, d), 8.03 (1H, s); m/z: ES+ [M+H]+ 485.

The5-fluoro-6-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-aminewas prepared as follows:

Preparation of1-(5-bromo-3-fluoropyridin-2-yl)-N-((1S,3R)-1-(5-bromo-3-fluoropyridin-2-yl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-yl)methanimine

5-Bromo-3-fluoropicolinaldehyde (1.09 g, 5.33 mmol) was added to(R)-3-(2-(((1-fluorocyclopropyl)methyl)amino)propyl)-2-methylaniline(630 mg, 2.67 mmol) and water (0.240 mL, 13.33 mmol) in acetic acid (12mL). The resulting solution was stirred at 70° C. for 2 hours. Thereaction mixture was concentrated and diluted with EtOAc (50 mL) andwashed sequentially with saturated aqueous NaHCO₃(50 mL). The organiclayer was dried with MgSO₄, filtered and evaporated to afford1-(5-bromo-3-fluoropyridin-2-yl)-N-((1S,3R)-1-(5-bromo-3-fluoropyridin-2-yl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-yl)methanimineas a crude product (1.53 g, 2.51 mmol). m/z: ES+ [M+H]+ 607.

Preparation of(1S,3R)-1-(5-bromo-3-fluoropyridin-2-yl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine

Hydroxylamine hydrochloride (0.174 g, 2.51 mmol) was added to1-(5-bromo-3-fluoropyridin-2-yl)-N-((1S,3R)-1-(5-bromo-3-fluoropyridin-2-yl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-yl)methanimine(1.53 g, 2.51 mmol) and potassium acetate (0.62 g, 6.27 mmol) in MeOH(12 mL). The resulting solution was stirred at 20° C. for 5 hours. Thereaction mixture was concentrated and diluted with DCM (75 mL) andwashed sequentially with NaOH (2M, 75 mL) and saturated aqueous sodiumchloride (50 mL). The organic layer was dried with MgSO₄, filtered andevaporated to afford crude product which was purified by flash silicachromatography, elution gradient 0 to 20% EtOAc in heptane. Productcontaining fractions were evaporated to dryness to afford(1S,3R)-1-(5-bromo-3-fluoropyridin-2-yl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(0.672 g, 63%) as a colourless gum. ¹H NMR (500 MHz, CDCl₃, 27° C.)0.45-0.61 (2H, m), 0.92-1.06 (2H, m), 1.07 (3H, d), 2.07 (3H, s), 2.51(1H, dd), 2.59 (1H, dd), 2.85 (1H, dd), 3.13 (1H, dd), 3.52 (2H, s),3.64-3.75 (1H, m), 5.35 (1H, s), 6.48 (2H, s), 7.52 (1H, dd), 8.36 (1H,dd); m/z: ES+ [M+H]+ 422/424.

Preparation of(6S,8R)-6-(5-bromo-3-fluoropyridin-2-yl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

(1S,3R)-1-(5-Bromo-3-fluoropyridin-2-yl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(0.672 g, 1.59 mmol) in propionic acid (6.63 mL) was cooled to −17° C.(dry ice/acetone). Sodium nitrite (0.110 g, 1.59 mmol) in water (1.33mL) was added dropwise and the reaction mixture stirred at −17° C. for30 minutes. The reaction mixture was diluted with ice-cold toluene (30mL), stirred at 0° C. for 15 minutes then at room temperature for 45minutes. The reaction mixture was washed with water (2×25 mL), thecombined aqueous phases washed with EtOAc (25 mL), the combined organicswere washed with saturated aqueous sodium chloride (50 mL), dried(MgSO₄), filtered and the filtrate evaporated to an orange-brown oil.The crude material was purified by flash silica chromatography, elutiongradient 0-50% EtOAc in heptane to afford(6S,8R)-6-(5-bromo-3-fluoropyridin-2-yl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.310 g, 45%) as an orange solid. ¹H NMR (500 MHz, CDCl₃, 27° C.)0.50-0.59 (2H, m), 1.03 (2H, dd), 1.13 (3H, d), 2.72 (1H, dd), 2.91 (1H,dd), 3.16 (1H, dd), 3.27 (1H, dd), 3.77-3.90 (1H, m), 5.50 (1H, s), 6.80(1H, d), 7.20 (1H, dd), 7.56 (1H, dd), 8.07 (1H, d), 8.36 (1H, dd); m/z:ES+ [M+H]+ 433.

Example 19 Preparation ofN-(4-((6S,8R)-7-(2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine

DMF (2 mL) and DIPEA (0.074 mL, 0.42 mmol) were added sequentially to aflask charged withN-(4-((6S,8R)-7-(2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)azetidin-3-amine(75 mg, 0.17 mmol). 1-Fluoro-3-iodopropane (31.9 mg, 0.17 mmol) in DMF(0.1 mL) was then added, and stirring was continued for 2 hours. Thereaction was stopped, diluted with saturated aqueous sodium chloride andthe compound was extracted in EtOAC (×3). The combined extracts werewashed with water and dried over sodium sulfate, filtered andconcentrated under reduced pressure to afford a film. This material waspurified by flash silica chromatography, eluting with 2 to 10% (methanolcontaining 1% ammonium hydroxide) in DCM to affordN-(4-((6S,8R)-7-(2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine(43 mg, 51%). ¹H NMR (400 MHz, DMSO-d₆, 27° C.) 1.00 (3H, d), 1.45-1.70(5H, m), 2.44 (2H, t), 2.55-2.66 (1H, m), 2.66-2.72 (2H, m), 2.79 (1H,dd), 2.87-3.01 (1H, m), 3.10 (1H, br dd), 3.38-3.53 (1H, m), 3.56-3.67(2H, m), 3.77 (3H, s), 3.85-3.97 (1H, m), 4.43 (2H, dt), 5.21 (1H, s),5.88 (1H, dd), 6.00 (1H, d), 6.16 (1H, d), 6.35 (1H, d), 6.64 (1H, d),7.17 (1H, d), 8.02 (1H, s), 12.92 (1H, d). m/z: (ES+), [M+H]+=502.

The starting materialN-(4-((6S,8R)-7-(2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)azetidin-3-aminewas prepared according to the procedures below.

Preparation of 2,2-difluoropropyl trifluoromethanesulfonate

Trifluoromethanesulfonic anhydride (3.29 ml, 19.5 mmol) was addeddropwise to a solution of 2,2-difluoropropan-1-ol (1.7 g, 18 mmol) inDCM (40 mL) at −10° C. (salt/ice bath). 2,6-Dimethylpyridine (2.5 mL, 21mmol) was then added, and the reaction was stirred for 1 hour underthese conditions. The reaction was then washed with water (×2), and theorganic layer was dried over Na₂SO₄, filtered and concentrated underreduced pressure (vacuum ˜200 mbars) to afford 2,2-difluoropropyltrifluoromethanesulfonate (2.1 g, 52%) as a red oil. ¹H NMR (400 MHz,CHLOROFORM-d, 27° C.) 4.48 (2H, t), 1.73 (3H, t).

Preparation of(R)-3-(2-((2,2-difluoropropyl)amino)propyl)-2-methylaniline

2,2-Difluoropropyl trifluoromethanesulfonate (1.68 g, 7.37 mmol) wasadded to a stirred solution of (R)-3-(2-aminopropyl)-2-methylaniline(1.1 g, 6.7 mmol) and DIPEA (1.52 ml, 8.71 mmol) in 1,4-dioxane (20 mL).The reaction was heated at 65° C. for 3 hours before being cooled toroom temperature and then concentrated under reduced pressure. Theresulting residue was dissolved in EtOAc (30 mL) and washed withsaturated aqueous NaHCO₃. The aqueous layer was extracted with EtOAc (20mL) and the combined organic layers were dried over Na₂SO₄, filtered andconcentrated under reduced pressure. The resulting red oil was purifiedby flash silica chromatography, elution gradient 30 to 90% ethyl acetatein hexanes, to afford(R)-3-(2-((2,2-difluoropropyl)amino)propyl)-2-methylaniline (1.02 g,63%) as a gum. ¹H NMR (500 MHz, DMSO-d₆, 27° C.) 0.90 (3H, d), 1.55 (3H,t), 1.75 (1H, br s), 1.97 (3H, s), 2.29-2.38 (1H, m), 2.69-2.76 (2H, m),2.86 (2H, br t), 4.69 (2H, s), 6.34 (1H, d), 6.47 (1H, d), 6.77 (1H, t).m/z: (ES+), [M+H]+=439.

Preparation of(1S,3R)-1-(4-bromo-2-methoxyphenyl)-2-(2,2-difluoropropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine

4-Bromo-2-methoxybenzaldehyde (1.07 g, 4.95 mmol) was added to asolution of (R)-3-(2-((2,2-difluoropropyl)amino)propyl)-2-methylaniline(0.600 g, 2.48 mmol) in AcOH (12 mL) and water (0.223 g, 12.4 mmol), andthe reaction was heated at 80° C. for 18 hours. After cooling, thevolatiles were concentrated under reduced pressure, and the resultingresidue was dissolved in EtOAc. The solution was neutralized by washingwith saturated aqueous NaHCO₃. The organic layer was combined withaqueous HCl (1N), and the biphasic mixture was stirred at roomtemperature for 30 minutes. The layers were then separated, and theorganic layer was washed with aqueous HCl (1N). The combined aqueouslayers were extracted with EtOAc and then basified by addition of solidK₂CO₃. The organic layer was then extracted with EtOAc (×2), and thecombined organic layers were dried over sodium sulfate, filtered, andconcentrated under reduced pressure. The resulting residue was purifiedby flash silica chromatography, elution gradient 10 to 60% ethyl acetatein hexanes, to afford(1S,3R)-1-(4-bromo-2-methoxyphenyl)-2-(2,2-difluoropropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(0.576 g, 53%) as a gum. ¹H NMR (400 MHz, DMSO-d₆, 27° C.) 0.95 (3H, d),1.54 (3H, t), 1.93 (3H, s), 2.27-2.43 (2H, m), 2.57-2.74 (1H, m),2.76-2.98 (1H, m), 3.19-3.26 (1H, m), 3.84 (3H, s), 4.63 (2H, s), 5.12(1H, s), 6.26 (1H, d), 6.38 (1H, d), 6.58 (1H, d), 6.94 (1H, dd), 7.16(1H, d). m/z: (ES+), [M+H]+=439.

Also isolated wasN-((1S,3R)-1-(4-bromo-2-methoxyphenyl)-2-(2,2-difluoropropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-yl)acetamide(0.081 g, 7%) as a gum.

Preparation of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

Sodium nitrite (0.049 g, 0.72 mmol) as a solution in water (0.750 mL)was added dropwise to a cooled solution of(1S,3R)-1-(4-bromo-2-methoxyphenyl)-2-(2,2-difluoropropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(0.30 g, 0.68 mmol) in propionic acid (2.5 mL) at −15° C., and thereaction was stirred for 1 hour under these conditions. Ice-cold EtOAc(10 mL) was added followed by saturated aqueous NaHCO₃ (10 mL) inportions. The layers were separated, and the the organic layer waswashed with saturated aqueous NaHCO₃. The combined aqueous layers wereextracted with EtOAc, and all organic layers were combined, dried overNa₂SO₄, filtered and concentrated under reduced pressure. The resultingresidue was purified by flash silica chromatography, elution gradient 20to 70% ethyl acetate in hexanes, to afford(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.23 g, 75%) as a gum. ¹H NMR (500 MHz, DMSO-d₆, 27° C.) 1.01 (3H, d),1.53 (3H, t), 2.84 (1H, dd), 2.96-3.09 (1H, m), 3.14 (1H, br dd),3.36-3.48 (1H, m), 3.89 (3H, s), 5.32 (1H, s), 6.64 (2H, app t), 6.94(1H, dd), 7.16-7.27 (2H, m), 8.06 (1H, s), 12.99 (1H, br s). 1H obscuredby DMSO. m/z: (ES+), [M+H]+=450.

Preparation of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoropropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineand(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoropropyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-2H-pyrazolo[4,3-f]isoquinoline

DCM (7 mL) was added to a flask charged with(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.3 g, 0.67 mmol) and 4-methylbenzenesulfonic acid hydrate (0.025 g,0.13 mmol). 3,4-Dihydro-2H-pyran (0.084 g, 1.0 mmol) was added, and thereaction was stirred at room temperature for 18 hours. The reaction waswashed with saturated aqueous sodium hydrogencarbonate, and the organiclayer was dried over Na₂SO₄, filtered, and concentrated under reducedpressure. The resulting residue was purified by flash chromatography,eluting with 5 to 40% ethyl acetate in hexanes, to afford(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoropropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(175 mg, 49%) as a gum. ¹H NMR (400 MHz, CHLOROFORM-d, 27° C.) 0.99-1.13(3H, br s), 1.48-1.88 (6H, m), 2.01-2.08 (1H, m), 2.08-2.18 (1H, m),2.48-2.70 (2H, m), 2.82 (1H, dt), 2.88-3.04 (1H, m), 3.20 (1H, br d),3.55-3.67 (1H, m), 3.63-3.73 (1H, m), 3.88 (3H, s), 3.94-3.98 (1H, m),5.42 (1H, br s), 5.63 (1H, dt), 6.64 (1H, dd), 6.73 (1H, dd), 6.86 (1H,dd), 7.03 (1H, t), 7.25-7.29 (1H, m), 7.98 (1H, d). m/z: (ES+),[M+H]+=534.

Also isolated was(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoropropyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-2H-pyrazolo[4,3-f]isoquinoline(68 mg, 19%) as a gum. ¹H NMR (400 MHz, CHLOROFORM-d, 27° C.) 1.02-1.07(3H, m), 1.42-1.53 (3H, m), 1.59-1.81 (3H, m), 2.03-2.09 (1H, m),2.18-2.25 (2H, m), 2.59 (1H, qd), 2.69 (1H, dd), 2.86-2.98 (1H, m),3.02-3.14 (1H, m), 3.46-3.57 (1H, m), 3.72-3.82 (1H, m), 3.87-3.90 (3H,m), 4.11-4.16 (1H, m), 5.31 (1H, s), 5.61-5.68 (1H, m), 6.60 (1H, dd),6.72 (1H, dd), 6.88 (1H, dt), 7.01-7.05 (1H, m), 7.36 (1H, d), 8.09 (1H,s). m/z: (ES+), [M+H]+=534.

Preparation of tert-butyl3-((4-((6S,8R)-7-(2,2-difluoropropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)amino)azetidine-1-carboxylate

Dioxane (3.5 mL) was added to a flask charged with Cs₂CO₃ (213 mg, 0.65mmol), tert-butyl 3-aminoazetidine-1-carboxylate (85 mg, 0.49 mmol), and(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoropropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(175 mg, 0.33 mmol). The reaction flask was evacuated and back filledwith nitrogen (×3). BrettPhos 3rd Generation Precatalyst (30 mg, 0.03mmol) was added, and the flask was again evacuated and back-filled withnitrogen (×3). The reaction was heated at 100° C. for 4 hours. Thereaction was cooled to room temperature, filtered, and concentratedunder reduced pressure. The resulting crude gum was purified by flashsilica chromatography, eluting with 15 to 70% ethyl acetate in hexanes,to afford tert-butyl3-((4-((6S,8R)-7-(2,2-difluoropropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)amino)azetidine-1-carboxylate(152 mg, 74%) as a film. ¹H NMR (400 MHz, CHLOROFORM-d, 27° C.) 1.05(3H, d), 1.42 (9H, s), 1.44-1.57 (3H, m), 1.57-1.63 (1H, m), 1.66-1.77(2H, m), 2.04-2.08 (1H, m), 2.09-2.18 (1H, m), 2.47-2.69 (2H, m), 2.79(1H, dt), 2.84-2.99 (1H, m), 3.16 (1H, br dd), 3.48-3.61 (1H, m), 3.69(3H, dt), 3.83 (3H, s), 3.94-4.04 (2H, m), 4.15 (1H, br s), 4.20-4.29(2H, m), 5.32 (1H, s), 5.63 (1H, dt), 5.86 (1H, dd), 6.07 (1H, t), 6.55(1H, dd), 6.78 (1H, dd), 7.20-7.23 (1H, m), 7.98 (1H, d). m/z: (ES+),[M+H]+=626.

Preparation ofN-(4-((6S,8R)-7-(2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)azetidin-3-amine

Methanol (1 mL) and then HCl in dioxane (4 M; 1 mL, 4 mmol) were addedsequentially to a flask charged with tert-butyl3-((4-((6S,8R)-7-(2,2-difluoropropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)amino)azetidine-1-carboxylate(140 mg, 0.22 mmol). After 2 hours, the reaction was concentrated underreduced pressure and the resulting residue was purified using an SCX-2cartage that had been pre-treated with methanol. The compound was elutedfirst with methanol and then ammonia in methanol (3N). Product fractionswere concentrated under reduced pressure to affordN-(4-((6S,8R)-7-(2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)azetidin-3-amine(91 mg, 92%) as a gum. The product was used in the next step withoutfurther purification. ¹H NMR (400 MHz, DMSO-d₆, 27° C.) 1.00 (3H, d),1.52 (3H, t), 2.55-2.59 (1H, m), 2.78 (1H, dd), 2.85-2.99 (1H, m), 3.09(1H, dd), 3.39-3.50 (1H, m), 3.55-3.66 (2H, m), 3.77 (3H, s), 4.03-4.14(1H, m), 5.21 (1H, s), 5.85 (1H, dd), 6.05 (1H, br d), 6.14 (1H, d),6.34 (1H, d), 6.63 (1H, d), 7.17 (1H, d), 8.02 (1H, s), 12.93 (1H, brs). 3H not observed. m/z: (ES+), [M+H]+=442.

Example 20 Preparation ofN-(3-ethoxy-44(6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine

DMF (2 mL) and DIPEA (0.050 mL, 0.28 mmol) were added sequentially to aflask charged withN-(3-ethoxy-4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-amine(0.051 g, 0.11 mmol). 1-Fluoro-3-iodopropane (0.021 g, 0.11 mmol) wasadded as a solution in DMF (0.2 mL), and after 2 hours, the reaction wasdiluted with saturated aqueous sodium hydrogencarbonate. The mixture wasextracted with ethyl acetate (×3), and the combined organic layers werewashed with water, dried over sodium sulfate, filtered, and concentratedunder reduced pressure. The resulting residue was purified by flashsilica chromatography, elution gradient 2 to 10% (methanol containing 1%ammonium hydroxide) in DCM. Product fractions were combined,concentrated under reduced pressure, and the resulting residue waspurified by flash silica chromatography using the above conditions toaffordN-(3-ethoxy-4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine(0.023 g, 40%). ¹H NMR (CHLOROFORM-d, 27° C.) 0.50 (2H, br d), 0.88-1.05(2H, m), 1.12 (3H, br d), 1.45 (3H, t), 1.75-1.87 (2H, m), 2.59-2.73(3H, m), 2.87-3.04 (3H, m), 3.04-3.15 (1H, m), 3.41 (1H, br dd),3.73-3.90 (3H, m), 3.93-4.18 (4H, m), 4.52 (2H, dt), 5.37 (1H, br s),5.99 (1H, dd), 6.13 (1H, br d), 6.80-6.89 (2H, m), 7.14 (1H, d),8.06-8.08 (1H, m), 10.02 (1H, br s). m/z: (ES+), [M+H]+=510.

Procedures used to prepareN-(3-ethoxy-4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-amineare described below.

Preparation of(1S,3R)-1-(4-bromo-2-ethoxyphenyl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine

4-Bromo-2-ethoxybenzaldehyde (0.775 g, 3.39 mmol) was added to asolution of(R)-3-(2-(((1-fluorocyclopropyl)methyl)amino)propyl)-2-methylaniline(0.400 g, 1.69 mmol) in AcOH (9 mL) and water (0.152 g, 8.46 mmol). Thereaction was heated at 80° C. for 18 hours. After cooling, the reactionwas concentrated under reduced pressure, and then the residue wasdissolved in EtOAc and washed with saturated aqueous NaHCO₃. The layerswere separated, and the organic phase was combined with aqueoushydrochloric acid (1N). After stirring the biphasic mixture for 30minutes, the layers were separated. The organic layer was washed withaqueous HCl (1N), and the combined aqueous layers were extracted withEtOAc (×2). The combined aqueous layers were then basified by additionof solid K₂CO₃ and extracted with EtOAc (×2). The organic extracts werecombined, dried over sodium sulfate, filtered, and concentrated underreduced pressure. The resulting residue was purified by flash silicachromatography, elution gradient 10 to 60% ethyl acetate in hexanes toafford(1S,3R)-1-(4-bromo-2-ethoxyphenyl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(0.48 g, 63%) as a solid. 1H NMR (400 MHz, DMSO-d₆, 27° C.) 0.40-0.59(2H, m), 0.83-0.93 (2H, m), 0.94 (3H, d), 1.35 (3H, t), 1.93-1.96 (3H,m), 2.45-2.53 (2H, m), 2.80-2.93 (2H, m), 3.57 (1H, br d), 4.14 (2H, q),4.58 (2H, s), 5.15 (1H, s), 6.26 (1H, d), 6.35 (1H, d), 6.86 (1H, d),6.95 (1H, dd), 7.15 (1H, d). m/z: (ES+), [M+H]+=447.

Preparation of(6S,8R)-6-(4-bromo-2-ethoxyphenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

Sodium nitrite (0.039 g, 0.56 mmol) as a solution in water (0.750 mL)was added dropwise to a cooled solution of(1S,3R)-1-(4-bromo-2-ethoxyphenyl)-2-((1-fluorocyclopropyl)methyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(0.240 g, 0.54 mmol) in propionic acid (3.0 mL) at −15° C., and thereaction was maintained under these conditions for 1 hour. Then ice-coldEtOAc (10 mL) was added, followed by saturated aqueous NaHCO₃ (15 mL) inportions. Once addition was complete and gas evolution ceased, thelayers were separated. The organic layer was washed with saturatedaqueous NaHCO₃ (×2), and the combined aqueous layers were extracted withEtOAc (×2). The combined organic layers were dried over sodium sulfate,filtered and concentrated under reduced pressure. The resulting residuewas purified by flash chromatography, elution gradient 20 to 60% ethylacetate in hexanes, to afford(6S,8R)-6-(4-bromo-2-ethoxyphenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.145 g, 59%) as a gum. ¹H NMR (500 MHz, DMSO-d₆, 27° C.) 0.45 (1H, brs), 0.57 (1H, br s), 0.83-0.98 (2H, m), 1.00 (3H, d), 1.37 (3H, t),2.54-2.63 (1H, m), 2.87-3.00 (2H, m), 3.27-3.32 (1H, m), 3.74 (1H, brd), 4.13-4.23 (2H, m), 5.32 (1H, s), 6.67 (1H, d), 6.94 (1H, s),6.96-6.99 (1H, m), 7.19 (1H, d), 7.22 (1H, d), 8.06 (1H, s), 12.96 (1H,s). m/z: (ES+), [M+H]+=458.

Preparation of(6S,8R)-6-(4-bromo-2-ethoxyphenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

DCM (3 mL) and 3,4-dihydro-2H-pyran (35.8 mg, 0.43 mmol) were added to aflask charged with(6S,8R)-6-(4-bromo-2-ethoxyphenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(130 mg, 0.28 mmol) and 4-methylbenzenesulfonic acid hydrate (10.79 mg,0.06 mmol). The reaction was stirred at room temperature for 18 hours.The reaction was washed with saturated aqueous sodium hydrogencarbonate, and the organic layer was dried over sodium sulfate,filtered, and concentrated under reduced pressure. The resulting browngum was purified by flash silica chromatography, elution gradient 5 to40% ethyl acetate in hexanes, to afford(6S,8R)-6-(4-bromo-2-ethoxyphenyl)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(147 mg, 96%) as a dry film. ¹H NMR (500 MHz, CHLOROFORM-d, 27° C.)0.42-0.51 (2H, m), 0.94-1.03 (2H, m), 1.07-1.12 (3H, m), 1.49 (3H, t),1.61-1.68 (1H, m), 1.73-1.79 (2H, m), 2.02-2.10 (1H, m), 2.12-2.20 (1H,m), 2.53-2.63 (2H, m), 2.93 (1H, dt), 3.10 (1H, ddd), 3.38-3.46 (1H, m),3.65-3.77 (1H, m), 3.83-3.92 (1H, m), 4.00-4.07 (1H, m), 4.12-4.19 (2H,m), 5.43 (1H, s), 5.66 (1H, ddd), 6.81 (1H, d), 6.89-6.93 (1H, m),6.95-7.00 (1H, m), 7.03-7.06 (1H, m), 7.25 (1H, dd), 8.03 (1H, s). m/z:(ES+), [M+H]+=542.

Preparation of tert-butyl3-((3-ethoxy-44(6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)amino)azetidine-1-carboxylate

Dioxane (2.7 mL) was added to a flask charged with(6S,8R)-6-(4-bromo-2-ethoxyphenyl)-74(1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(140 mg, 0.26 mmol), Cs₂CO₃ (168 mg, 0.52 mmol) and tert-butyl3-aminoazetidine-1-carboxylate (67 mg, 0.39 mmol). The reaction flaskwas evacuated and filled with N₂ (×3). BrettPhos 3^(rd) GenerationPrecatalyst (23 mg, 0.030 mmol) was added, and the flask was evacuatedand filled with nitrogen (×3). The reaction was heated at 90° C. for 1hour and then at 90° C. for 44 hours. The reaction was cooled to roomtemperature, filtered, and concentrated under reduced pressure. Theresulting gum was purified by flash chromatography, elution gradient 30to 100% ethyl acetate in hexanes, to afford tert-butyl3-((3-ethoxy-4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)amino)azetidine-1-carboxylate(86 mg, 53%) as a gum. ¹H NMR (500 MHz, CHLOROFORM-d, 27° C.) 0.39-0.49(2H, m), 0.87-0.95 (2H, m), 1.06 (3H, d), 1.38-1.45 (12H, m), 1.57-1.62(1H, m), 1.68-1.74 (2H, m), 1.98-2.06 (1H, m), 2.07-2.15 (1H, m),2.49-2.63 (2H, m), 2.85 (1H, dt), 3.05 (1H, br t), 3.29-3.35 (1H, m),3.63-3.73 (3H, m), 3.75-3.83 (1H, m), 3.92 (1H, br d), 3.99 (1H, br d),4.05 (2H, q), 4.14 (1H, br dd), 4.18-4.26 (2H, m), 5.33 (1H, s),5.58-5.63 (1H, m), 5.90 (1H, br d), 6.02-6.06 (1H, m), 6.76 (1H, dd),6.80 (1H, d), 7.19 (1H, d), 7.97 (1H, s). m/z: (ES+), [M+H]+=634.

Preparation ofN-(3-ethoxy-4-((6S,8R)-7-((l-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-amine

Methanol (0.5 mL) was added to a flask charged with tert-butyl3-((3-ethoxy-4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)amino)azetidine-1-carboxylate(0.080 g, 0.13 mmol). Hydrochloric acid in dioxane (4 M; 0.5 mL, 2 mmol)was added, and stirring was continued for 2 hours. The reaction was thenconcentrated under reduced pressure, and the resulting residue waspurified using an SCX-2 cartridge that had been pre-treated withmethanol, eluting first with methanol and then with ammonia in methanol(3N), to affordN-(3-ethoxy-4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-amine(0.057 g, 99%) as a solid. ¹H NMR (500 MHz, CHLOROFORM-d, 27° C.)0.44-0.55 (2H, m), 0.90-1.00 (2H, m), 1.12 (3H, d), 1.45 (3H, t),2.61-2.71 (1H, m), 2.92 (1H, br dd), 3.10 (1H, br dd), 3.40 (1H, br dd),3.51-3.58 (2H, m), 3.83-3.90 (1H, m), 3.91-4.02 (2H, m), 4.03-4.16 (3H,m), 4.36 (1H, sxt), 5.38 (1H, s), 5.98 (1H, dd), 6.12 (1H, br d),6.77-6.89 (2H, m), 7.13 (1H, d), 8.07 (1H, s), 10.09 (1H, br s). One Hnot observed and likely obscured by water peak. m/z (ES+), [M+H]+=450.

Example 21 Preparation ofN-(4-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine

Hydrochloric acid in dioxane (4 M; 1.23 mL, 5.2 mmol) was addeddropwisely to a stirred solution of tert-butyl3-(4-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-2H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenylamino)azetidine-1-carboxylate(316 mg, 0.52 mmol) in MeOH (5 mL). The mixture was stirred at roomtemperature for 18 hours. The reaction was concentrated under reducedpressure, and the resulting residue was dissolved in MeOH. Excesstetraalkylammonium carbonate macroporous resin (Aldrich; 18-50 mesh;2.5-3.5 mmol/g N loading) was added, and the mixture was stirred at roomtemperature for 5 minutes. The mixture was filtered, and the filtratewas dried over sodium sulfate, filtered, and concentrated under reducedpressure to afford crudeN-(4-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)azetidin-3-amineas a gum (220 mg). This material was directly used in next step withoutfurther purification. m/z: (ES+), [M+H]+=428.

A mixture ofN-(4-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)azetidin-3-amine(220 mg, 0.51 mmol), 1-fluoro-3-iodopropane (106 mg, 0.57 mmol), andDIPEA (0.270 mL, 1.54 mmol) in DMF (5 mL) was stirred at roomtemperature for 18 hours. The reaction was diluted with DCM and washedwith saturated aqueous ammonium chloride. The aqueous layer wasextracted with DCM, and the combined organic layers were dried oversodium sulfate, filtered, and concentrated under reduced pressure. Theresulting residue was purified by preparative HPLC (Xbridge C18 column,19 mm×150 mm; 5 μm; flow rate: 20 mL/min), eluting with 40 to 80%acetonitrile in water containing 0.2% ammonium hydroxide. Productfractions were concentrated under reduced pressure, and the resultingresidue was purified by preparative SFC (2-ethylpyridine column, 19mm×150 mm, 5 μm; flow rate: 75 mL/min; column temperature: 40° C.;outlet pressure: 100 bar), eluting with 15% methanol containing 0.2%ammonium hydroxide in carbon dioxide, to affordN-(4-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine(82 mg, 33% two steps) as a white solid. ¹H NMR (300 MHz, DMSO-d₆, 27°C.) 1.03 (3H, d), 1.55-1.74 (2H, m), 2.41-2.47 (2H, m), 2.53-2.83 (4H,m), 2.86-3.04 (1H, m), 3.10 (1H, br dd), 3.32-3.43 (1H, m), 3.58-3.66(2H, m), 3.82 (3H, s), 3.86-3.98 (1H, m), 4.45 (2H, dt), 5.21 (1H, s),5.89 (1H, t), 5.90 (1H, dd), 6.01 (1H, d), 6.19 (1H, d), 6.35 (1H, d),6.67 (1H, d), 7.19 (1H, d), 8.03 (1H, s), 12.92-12.96 (1H, m). m/z: ES+[M+H]+ 488.

The starting material tert-butyl3-((4-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)amino)azetidine-1-carboxylatewas prepared according to the following procedures.

Preparation of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineand(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoroethyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-2H-pyrazolo[4,3-f]isoquinoline

A microwave vial was charged with(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(986 mg, 2.26 mmol), 4-methylbenzenesulfonic acid hydrate (43 mg, 0.23mmol), 3,4-dihydro-2H-pyran (0.31 mL, 3.4 mmol) and DCM. The reactionwas heated at 80° C. under microwave conditions (300 W) for 20 minutes.The reaction was then cooled and diluted with DCM before being washedwith saturated aqueous sodium hydrogencarbonate. The layers wereseparated, and the organic layer was dried over sodium sulfate,filtered, and concentrated under reduced pressure. The resulting residuewas purified by flash silica chromatography, elution gradient 0 to 70%EtOAc in hexanes, to afford faster eluting(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(457 mg, 39%) as an orange solid. ¹H NMR (300 MHz, CHLOROFORM-d, 27° C.)1.04-1.22 (3H, m), 1.59-1.86 (3H, m), 2.04-2.23 (2H, m), 2.53-3.31 (5H,m), 3.40-3.57 (1H, m), 3.68-3.79 (1H, m), 3.97 (3H, s), 4.00-4.09 (1H,m), 5.41 (1H, br s), 5.69 (1H, br s), 5.66-5.73 (1H, m), 6.67 (1H, dd),6.80 (1H, dd), 6.92 (1H, dd), 7.06-7.12 (1H, m), 7.29-7.37 (1H, m), 8.03(1H, s). m/z: ES+ [M+H]+ 612. Also isolated was slower eluting(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoroethyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-2H-pyrazolo[4,3-f]isoquinoline(627 mg, 57%) as a beige powder. ¹H NMR (300 MHz, CHLOROFORM-d, 27° C.)1.12 (3H, br d), 1.64-1.87 (3H, m), 2.03-2.17 (1H, m), 2.18-2.29 (2H,m), 2.61-2.83 (2H, m), 2.89-3.15 (2H, m), 3.43 (1H, br s), 3.75-3.86(1H, m), 3.95 (3H, s), 4.13-4.20 (1H, m), 5.32 (1H, s), 5.79 (1H, br t),5.66-5.73 (1H, m), 6.63-6.77 (2H, m), 6.93 (1H, d), 7.09 (1H, d), 7.43(1H, d), 8.14 (1H, s). m/z: ES+ [M+H]+ 612.

Preparation of tert-butyl3-(4-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-2H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenylamino)azetidine-1-carboxylate

A microwave vial was charged with(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoroethyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-2H-pyrazolo[4,3-f]isoquinoline(310 mg, 0.60 mmol), tert-butyl 3-aminoazetidine-1-carboxylate (154 mg,0.89 mmol), Cs₂CO₃ (388 mg, 1.19 mmol), and BrettPhos 3^(rd) GenerationPrecatalyst (54.0 mg, 0.06 mmol). The vial was degassed and filled withnitrogen (×2). The vial was degassed again and re-filled with1,4-dioxane (6 mL) and nitrogen. The vial was degassed again andre-filled with nitrogen. The mixture was heated under microwaveconditions (300 W, 110° C.) for 3.5 hours. The reaction was diluted withDCM and washed with saturated aqueous sodium hydrogencarboante. Theorganic layer was dried over sodium sulfate, filtered, and concentratedunder reduced pressure. The resulting residue was purified by flashsilica chromatography, elution gradient 20 to 60% EtOAc in hexanes, toafford tert-butyl3-(4-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-2-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-2H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenylamino)azetidine-1-carboxylate(322 mg, 88%) as an off-white solid. ¹H NMR (300 MHz, CHLOROFORM-d, 27°C.) 1.08 (3H, br d), 1.39-1.43 (9H, m), 1.58-1.81 (3H, m), 2.06 (1H, brdd), 2.13-2.25 (2H, m), 2.58-2.71 (1H, m), 2.82-3.11 (2H, m), 3.34-3.50(1H, m), 3.66-3.81 (3H, m), 3.86 (3H, s), 3.90-4.01 (1H, m), 4.08-4.20(3H, m), 4.19-4.29 (2H, m), 5.22 (1H, s), 5.76 (1H, t), 5.64 (1H, t),5.88 (1H, br d), 6.09 (1H, d), 6.59 (1H, br t), 6.67 (1H, d), 7.37 (1H,br d), 8.08 (1H, s). m/z: ES+ [M+H]+ 612.

Example 22 Preparation of(6S,8R)-7-(2,2-difluoroethyl)-6-(4-((1-(3-fluoropropyl)azetidin-3-yl)oxy)-2-methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

HCl in dioxane (4 M; 1.02 mL, 4.08 mmol) was added dropwisely to astirred solution of tert-butyl3-(4-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenoxy)azetidine-1-carboxylate(250 mg, 0.41 mmol) in MeOH (4 mL). The mixture was stirred at roomtemperature for 18 hours. The reaction was then concentrated underreduced pressure, and the resulting residue was dissolved with MeOH.Excess tetraalkylammonium carbonate macroporous resin (Aldrich; 18-50mesh; 2.5-3.5 mmol/g N loading) was added, and the mixture was stirredat room temperature for 5 minutes before being dried over sodiumsulfate, filtered, and concentrated under reduced pressure to affordcrude(6S,8R)-6-(4-(azetidin-3-yloxy)-2-methoxyphenyl)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(179 mg), which was directly used in next step without furtherpurification. m/z: ES+ [M+H]+ 429.

A mixture of(6S,8R)-6-(4-(azetidin-3-yloxy)-2-methoxyphenyl)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(178 mg, 0.42 mmol), 1-fluoro-3-iodopropane (94 mg, 0.50 mmol), andDIPEA (0.218 mL, 1.25 mmol) in DMF (4 mL) was stirred at roomtemperature for 18 hours. The reaction was then diluted with DCM andwashed with saturated aqueous ammonium chloride. The layers wereseparated, and the aqueous layer was extracted with DCM. The combinedorganic layers were dried over sodium sulfate, filtered, andconcentrated under reduced pressure. The resulting residue was purifiedby preparative SFC (2-ethylpyridine column; 19 mm×150 mm; 5 μm; flowrate: 75 mL/min; column temperature: 40° C.; outlet pressure: 100 bar),eluting with 15% (methanol containing 0.2% ammonium hydroxide) in carbondioxide to afford(6S,8R)-7-(2,2-difluoroethyl)-6-(4-((1-(3-fluoropropyl)azetidin-3-yl)oxy)-2-methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(70 mg, 35% over two steps) as a white solid. ¹H NMR (300 MHz, DMSO-d₆,27° C.) 1.02 (3H, d), 1.61-1.84 (2H, m), 2.55-2.84 (3H, m), 2.90-3.13(2H, m), 3.18-3.43 (4H, m), 3.87 (3H, s), 3.89-4.00 (2H, m), 4.45 (2H,dt), 4.81 (1H, quin), 5.28 (1H, s), 5.74-6.16 (1H, m), 6.19 (1H, dd),6.49-6.54 (2H, m), 6.66 (1H, d), 7.21 (1H, d), 8.04 (1H, s), 12.98 (1H,s). m/z: ES+ [M+H]+ 489.

Procedures used to prepare the starting material tert-butyl3-(4-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenoxy)azetidine-1-carboxylateare described below.

Preparation of tert-butyl3-(4-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenoxy)azetidine-1-carboxylate

A microwave vial was charged with(6S,8R)-6-(4-bromo-2-methoxyphenyl)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(259 mg, 0.50 mmol), tert-butyl 3-hydroxyazetidine-1-carboxylate (259mg, 1.49 mmol), cesium carbonate (324 mg, 1.00 mmol), and RockPhos 3rdGeneration Precatalyst (42 mg, 0.050 mmol). The vial was evacuated andre-filled with nitrogen (×2). Then the vial was evacuated and re-filledwith toluene (3 mL). The vial was evacuated and re-filled with nitrogen.The mixture was heated at 110° C. for 4 hours. The reaction was dilutedwith DCM and washed with saturated aqueous sodium hydrogencarbonate. Theorganic layer was dried over sodium sulfate, filtered, and concentratedunder reduced pressure. The resulting residue was purified by flashsilica chromatography, elution gradient 0 to 60% EtOAc in hexanes toafford tert-butyl3-(4-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenoxy)azetidine-1-carboxylate(220 mg, 72%) as a white foam solid. ¹H NMR (400 MHz, DMSO-d₆, 100° C.)1.03-1.09 (3H, m), 1.40 (9H, s), 1.56-1.64 (2H, m), 1.67-1.84 (1H, m),1.93-2.01 (1H, m), 2.02-2.12 (1H, m), 2.34-2.46 (1H, m), 2.59-2.74 (1H,m), 2.84 (1H, dt), 2.93-3.06 (1H, m), 3.13-3.23 (1H, m), 3.40-3.50 (1H,m), 3.65-3.74 (1H, m), 3.76-3.84 (2H, m), 3.84-3.91 (4H, m), 4.21-4.28(2H, m), 4.97 (1H, tt), 5.33 (1H, s), 5.67-6.00 (1H, m), 5.74 (1H, dd),6.23 (1H, ddd), 6.54 (1H, d), 6.66 (1H, t), 6.73 (1H, d), 7.35 (1H, d),8.04 (1H, s). m/z: ES+ [M+H]+ 613.

Examples 23 and 24 Preparation of of individual diastereoisomers of3-((6S,8R)-6-(2,6-difluoro-4-(1-(3-fluoropropyl)azetidin-3-ylamino)phenyl)-8-methyl-8,9-dihydro-3H-pyrazolo[4,3-f]isoquinolin-7(6H)-yl)-2-fluoro-2-methylpropan-1-ol

Tetrabutylammonium fluoride in THF (1 M; 0.114 mL, 0.11 mmol) was addeddropwise via syringe to a stirred solution ofN-(4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)-1-(3-fluoropropyl)azetidin-3-amine(82 mg, 0.08 mmol) in THF (0.75 mL). After 2.5 hours additionaltetrabutylammonium fluoride in THF (1 M; 0.08 mL, 0.08 mmol) was addedand the reaction was stirred for a further 18 hours. The reaction wasdiluted with DCM and washed successively with saturated aqueous NaHCO₃and saturated aqueous sodium chloride, dried over MgSO₄, filtered andconcentrated under reduced pressure. The resulting residue was purifiedby flash silica chromatography, elution gradient 0.5 to 10% MeOH in DCM.Product fractions were concentrated under reduced pressure and theresulting residue was further purified by preparative HPLC (WatersXBridge Prep C18 OBD column, 5 μm silica, 19 mm diameter, 150 mmlength), using an elution gradient of 45 to 85% acetonitrile in (watercontaining 0.2% NH₄OH). Product fractions were concentrated underreduced pressure to afford first eluting isomer 1 (2.7 mg, 7%) andsecond eluting isomer (2.1 mg, 5%) of3-((6S,8R)-6-(2,6-difluoro-4-(1-(3-fluoropropyl)azetidin-3-ylamino)phenyl)-8-methyl-8,9-dihydro-3H-pyrazolo[4,3-f]isoquinolin-7(6H)-yl)-2-fluoro-2-methylpropan-1-olas pale yellow films.

Isomer 1: ¹H NMR (500 MHz, CD₂Cl₂, 27° C.) 1.13 (3H, d), 1.17 (3H, d),1.66-1.78 (3H, m), 2.54 (2H, t), 2.69 (1H, dd), 2.81-2.94 (3H, m), 3.00(1H, dd), 3.31 (1H, dd), 3.51-3.59 (1H, m), 3.60-3.71 (4H, m), 3.95-4.03(1H, m), 4.38 (1H, br s), 4.46 (2H, dt), 5.26 (1H, s), 6.01 (2H, br d),6.84 (1H, d), 7.21 (1H, d), 8.03 (1H, s), 10.19 (1H, br s). m/z:ES+[M+H]+ 520.

Isomer 2: ¹H NMR (500 MHz, CD₂Cl₂, 27° C.) 1.05 (3H, d), 1.09 (3H, d),1.65-1.80 (2H, m), 2.54 (2H, t), 2.62 (1H, dd), 2.83-2.88 (2H, m),2.91-3.00 (1H, m), 3.13-3.24 (1H, m), 3.33-3.46 (2H, m), 3.48-3.58 (1H,m), 3.66 (2H, q), 3.96-4.04 (2H, m), 4.40 (1H, dt), 4.41-4.53 (2H, m),4.64 (1H, br d), 5.03 (1H, s), 6.06 (2H, br d), 6.76 (1H, d), 7.19 (1H,d), 8.03 (1H, s), 10.19 (1H, br s). m/z: ES+ [M+H]+ 520.

Procedures used to prepare the starting materialN-(4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)-1-(3-fluoropropyl)azetidin-3-amineare described below.

Preparation of dimethyl 2-fluoro-2-methyl-propanedioate

Sodium hydride (60% dispersion in mineral oil; 2.93 g, 73.3 mmol) wasadded to a solution of dimethyl 2-fluoromalonate (10.0 g, 66.6 mmol) inTHF (218 mL) with vigorous stirring. After 30 minutes iodomethane (4.56mL, 73.3 mmol) was added. The reaction was stirred for a further 3hours. The reaction was quenched with water and then extracted withEtOAc (4×100 mL). The combined organic layers were dried over Na₂SO₄,filtered, and concentrated under reduced pressure to afford dimethyl2-fluoro-2-methyl-propanedioate (8.4 g, 77%) as an orange oil. ¹H NMR(300 MHz, DMSO-d₆, 27° C.) 1.72 (3H, d), 3.77 (6H, s).

Preparation of 2-fluoro-2-methylpropane-1,3-diol

Lithium aluminium hydride (4.50 g, 112.6 mmol) was added portion-wise toa stirred solution of dimethyl 2-fluoro-2-methyl-propanedioate (8.40 g,51.2 mmol) in THF (205 mL) at 0° C. The reaction was warmed to roomtemperature and stirred under these conditions for 1 hour. The reactionmixture was then cooled to 0° C. and cautiously quenched by sequentialdropwise addition of water (5.85 mL), 15 wt % aqueous NaOH (5.85 mL),and water (18 mL). The resulting gelatinous suspension was stirredrapidly for 1 hour. The precipitate was removed by filtration, and thefiltrate was concentrated under reduced pressure. The resulting residuewas dissolved in a mixture of CHCl₃/IPA (3:1), dried over Na₂SO₄,filtered, and concentrated under reduced pressure to afford2-fluoro-2-methylpropane-1,3-diol (3.28 g, 46%) as an orange oil. Thisoil was used in the next step without further purification. ¹H NMR (300MHz, DMSO-d₆, 27° C.) 1.18 (3H, d), 3.42 (4H, dd), 4.80 (2H, t).

Preparation of3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropan-1-ol

Sodium hydride (60% dispersion in mineral oil; 1.1 g, 27.5 mmol) wasadded to a stirred solution of 2-fluoro-2-methylpropane-1,3-diol (2.7 g,25 mmol) in THF (93 mL) at 0° C. and the reaction was stirred for 1hour. Tert-butylchlorodiphenylsilane (6.5 mL, 25 mmol) was added, andthe reaction was stirred for an additional 1 hour. The reaction wasquenched with water, the layers were separated, and the aqueous layerwas extracted with EtOAc (2×100 mL). The combined organic layers weredried over Na₂SO₄, filtered, and adsorbed onto silica gel under reducedpressure. Purification by flash silica chromatography, elution gradient0 to 50% EtOAc in hexanes, afforded3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropan-1-ol (4.4 g,51%) as a clear gum. ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 1.00 (9H, s),1.27 (3H, d), 3.48 (1H, dd), 3.53 (1H, dd), 3.66 (1H, d), 3.73 (1H, brs), 4.93 (1H, t), 7.40-7.48 (6H, m), 7.59-7.66 (4H, m). m/z: ES+[M+H]+347.

Preparation of 3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyltrifluoromethanesulfonate

Trifluoromethanesulfonic anhydride (1.3 mL, 7.6 mmol) was added dropwiseto a stirred solution of3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropan-1-ol (2.2 g,6.35 mmol) and 2,6-dimethylpyridine (1.28 mL, 7.6 mmol) in DCM (22 mL)at −10° C. (salt/ice bath). The reaction was maintained under theseconditions for 1.5 hours. The reaction was then diluted with DCM (100mL) and washed successively with aqueous HCl (1N), saturated aqueousNaHCO₃, and saturated aqueous NaCl. The organic layer was dried overNa₂SO₄, filtered, and concentrated under reduced pressure to afford3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyltrifluoromethanesulfonate (3.2 g) as a red oil. This oil was used in thenext step without further purification.

Preparation of a diastereoisomeric mixture of3-((2R)-2-((3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)amino)propyl)-2-methylaniline

3-((Tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyltrifluoromethanesulfonate (3.04 g, 6.35 mmol) was added to a solution of(R)-3-(2-aminopropyl)-2-methylaniline (1.04 g, 6.35 mmol) anddiisopropylethylamine (1.65 mL, 9.53 mmol) in 1,4-dioxane (24 mL). Thereaction was heated at 85° C. for 18 hours. After cooling, the reactionwas diluted with DCM (250 mL) and washed with water. The aqueous layerwas extracted with DCM (2×100 mL), and the combined organics were driedover Na₂SO₄, filtered, and concentrated under reduced pressure. Theresulting residue was purified by flash silica chromatography, elutiongradient 0 to 25% MeOH in DCM. Product fractions were concentrated underreduced pressure to afford3-((2R)-2-((3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)amino)propyl)-2-methylaniline(2.10 g, 95%) as a gum. ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 0.90 (3H, dd),1.01 (9H, d), 1.29 (3H, dd), 1.41 (1H, br s), 1.97 (3H, d), 2.29-2.43(1H, m), 2.65-2.91 (4H, m), 3.62-3.82 (2H, m), 4.67 (2H, s), 6.28-6.36(1H, m), 6.47 (1H, dd), 6.69-6.81 (1H, m), 7.40-7.52 (6H, m), 7.61-7.69(4H, m). m/z: ES+ [M+H]+ 494.

Preparation of a diastereoisomeric mixture of(1S,3R)-1-(4-bromo-2,6-difluorophenyl)-2-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine

4-bromo-2,6-difluorobenzaldehyde (628 mg, 2.84 mmol) was added to astirred solution of3-((2R)-2-((3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)amino)propyl)-2-methylaniline(700 mg, 1.42 mmol) and water (0.128 mL, 7.10 mmol) in a mixture ofacetic acid (10 mL) and toluene (4 mL). The reaction was heated at 90°C. for 18 hours before being heated at reflux conditions for another 4hours. The reaction was allowed to cool and was concentrated underreduced pressure. The resulting residue was dissolved in DCM and washedwith saturated aqueous NaHCO₃. The aqueous phase was extracted with DCM(20 mL) and the combined organic layers were concentrated under reducedpressure. The residue was dissolved in a mixture of MeOH/DCM (5:1, 18mL) and then hydroxylamine hydrochloride (148 mg, 2.13 mmol) and sodiumacetate (233 mg, 2.84 mmol) were added. The mixture was stirred at 35°C. for 5 minutes and then concentrated under reduced pressure. Theresulting residue was dissolved in EtOAc and washed sequentially withsaturated aqueous NaHCO₃ and saturated aqueous NaCl before being driedover MgSO₄, filtered and concentrated under reduced pressure. Theresulting residue was purified by flash silica chromatography, elutiongradient 0 to 35% EtOAc in hexanes to afford(1S,3R)-1-(4-bromo-2,6-difluorophenyl)-2-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(339 mg, 34%) as a pale yellow film. ¹H NMR (500 MHz, CDCl₃, 27° C.)1.05 (9H, d), 1.19 (3H, dd), 2.08 (3H, s), 2.35 (1H, dd), 2.47-2.67 (2H,m), 2.84-3.01 (1H, m), 3.02-3.14 (1H, m), 3.22 (1H, dd), 3.40-3.63 (3H,m), 3.66-3.79 (1H, m), 3.92 (1H, dd), 5.15 (1H, d), 6.38-6.47 (2H, m),6.83 (1H, d), 6.93-7.00 (1H, m), 7.38-7.49 (6H, m), 7.59-7.71 (4H, m),8.69 (1H, s). m/z: ES+ [M+H]+ 695.

Preparation of a diastereoisomeric mixture of(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

Sodium nitrite (33.6 mg, 0.49 mmol) as a solution in water (0.400 mL)was added dropwise to a cooled solution of(1S,3R)-1-(4-bromo-2,6-difluorophenyl)-2-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(339 mg, 0.49 mmol) in propionic acid (4 mL) at −20° C., and thereaction was stirred for 30 minutes under these conditions. Ice-coldEtOAc (20 mL) was added followed by portion-wise addition of saturatedaqueous NaHCO₃ (5 mL).

The biphasic mixture was stirred vigorously and neutralized by slowaddition of solid Na₂CO₃. The phases were separated, and the organiclayers were washed with saturated aqueous NaHCO₃ (2×30 mL) and saturatedaqueous NaCl (30 mL) before being dried over MgSO₄, filtered, andconcentrated under reduced pressure. The resulting residue was purifiedby flash silica chromatography, elution gradient 0 to 40% EtOAc inhexanes, to afford(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(206 mg, 60%) as a pale yellow solid. ¹H NMR (500 MHz, CDCl₃, 27° C.)0.98 (4.5H, s), 1.00 (1.5H, d), 1.02 (1.5H, d), 1.04 (4.5H, s), 1.15(1.5H, d), 1.23 (1.5H, d), 2.30-2.47 (0.5H, m), 2.60 (0.5H, dd), 2.81(0.5H, dd), 2.89 (0.5H, dd), 3.02 (0.5H, dd), 3.06-3.21 (1H, m),3.22-3.33 (0.5H, m), 3.37-3.51 (1H, m), 3.51-3.60 (0.5H, m), 3.61-3.72(0.5H, m), 3.73-3.83 (0.5H, m), 3.89 (0.5H, dd), 5.22 (0.5H, s), 5.27(0.5H, s), 6.70 (1H, dd), 6.81 (1H, br d), 6.91-7.00 (1H, m), 7.14 (1H,t), 7.33-7.47 (6H, m), 7.55-7.67 (4H, m), 8.07 (1H, br d). Indazole NHnot observed. m/z: ES+ [M+H]+ 706.

Preparation of a diastereoisomeric mixture of(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

3,4-Dihydro-2H-pyran (0.129 mL, 1.41 mmol) and para-toluenesulfonic acidmonohydrate (2.7 mg, 0.01 mmol) were added to a stirred solution of(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(200 mg, 0.28 mmol) in DCM (2 mL). The reaction was heated at 45° C. for21 hours. The reaction was allowed to cool and then diluted with DCM,washed with saturated aqueous NaHCO₃, dried over MgSO₄, filtered, andconcentrated under reduced pressure. The resulting residue was purifiedby flash silica chromatography, elution gradient 5 to 30% EtOAc inhexanes, to afford(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(176 mg, 79%) as a colorless film. m/z: ES+ [M+H]+ 790.

Preparation of a diastereoisomeric mixture of tert-butyl3-((4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)amino)azetidine-1-carboxylate

A vial was charged with a stirrer bar,(6S,8R)-6-(4-bromo-2,6-difluorophenyl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(126 mg, 0.16 mmol), tert-butyl 3-aminoazetidine-1-carboxylate (41 mg,0.24 mmol), Pd₂dba₃ (9.3 mg, 0.01 mmol),4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (Xantphos) (16 mg, 0.02mmol), and cesium carbonate (156 mg, 0.48 mmol). The vial was sealed,evacuated and backfilled with nitrogen (3×) prior to the addition of1,4-dioxane (1 mL) via syringe. The mixture was stirred at ambienttemperature for 2 minutes and then heated at 90° C. for 16 hours. Themixture was allowed to cool and then diluted with EtOAc, filteredthrough diatomaceous earth, and concentrated under reduced pressure. Theresulting residue was purified by flash silica chromatography, elutiongradient 0 to 40% EtOAc in hexanes, to afford tert-butyl3-((4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)amino)azetidine-1-carboxylate(95 mg, 68%) as a pale yellow solid. m/z: ES+ [M+H]+ 883.

Preparation of a diastereoisomeric mixture ofN-(4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)azetidin-3-amine

Formic acid (0.5 mL, 13.04 mmol) was added to tert-butyl3-((4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-j]isoquinolin-6-yl)-3,5-difluorophenyl)amino)azetidine-1-carboxylate(53 mg, 0.06 mmol) and the resulting solution was stirred at ambienttemperature for 18 hours. The reaction was concentrated under reducedpressure, and the resulting residue was dissolved in THF (0.5 mL) andtreated with aqueous NaOH (5N; 0.081 mL, 2.40 mmol). The mixture wasstirred at 30° C. for 20 hours and then maintained at 40° C. for 4hours. Additional aqueous NaOH (5N; 0.081 mL, 2.40 mmol) was added, andthe reaction stirred at 40° C. for 46 hours before being heated at 55°C. for 5 hours. The reaction mixture was allowed to cool to ambienttemperature. Meanwhile, in a separate vial, HCl in dioxane (4N; 0.38 mL,1.5 mmol) was added to a stirred solution of tert-butyl3-((4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)amino)azetidine-1-carboxylate(89 mg, 0.10 mmol) in MeOH (0.5 mL). The reaction was stirred at roomtemperature for 5 hours and then concentrated under reduced pressure.The resulting residue was combined with the previous reaction (as amixture in aqueous NaOH (5N) and THF). The new mixture was diluted withDCM (5 mL), the phases were separated, and the aqueous phase wasextracted with DCM (2×5 mL). The combined organic layers were dried overMgSO₄, filtered and concentrated under reduced pressure to affordN-(4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)azetidin-3-amine(177 mg) as a pale yellow solid, which was used in the next step withoutfurther purification. m/z: ES+[M+H]+ 698.

Preparation of a diastereoisomeric mixture ofN-(4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)-1-(3-fluoropropyl)azetidin-3-amine

1-Fluoro-3-iodopropane (0.026 mL, 0.16 mmol) was added to a stirredsolution ofN-(4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)azetidin-3-amine(115 mg, 0.16 mmol) and diisopropylethylamine (0.058 mL, 0.33 mmol) inNMP (0.75 mL). The reaction was stirred for 16 hours at room temperaturebefore being diluted with EtOAc and washed with saturated aqueousNaHCO₃. The layers were separated and the aqueous layer was extractedwith EtOAc. The combined organic layers were washed with saturatedaqueous NaCl (2×5 mL), dried over MgSO₄, filtered, and concentratedunder reduced pressure. The resulting residue was purified by flashsilica chromatography, elution gradient 1 to 20% MeOH in DCM to affordN-(4-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2-fluoro-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)-1-(3-fluoropropyl)azetidin-3-amine(82 mg, 66%) as a pale yellow film. m/z: ES+ [M+H]+ 758.

Example 25 Preparation of6-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine

HCl in dioxane (4N; 0.86 mL, 3.4 mmol) was added dropwise to a solutionof tert-butyl3-((6-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-yl)amino)azetidine-1-carboxylate(0.20 g, 0.34 mmol) in MeOH (2.5 mL), and the reaction was stirred atroom temperature for 2 hours. The reaction was concentrated underreduced pressure to affordN-(azetidin-3-yl)-6-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-aminehydrochloride (200 mg) as a solid. m/z: (ES+), [M+H]+ 399.

DMF (0.5 mL) and 1-fluoro-3-iodopropane (0.036 mL, 0.34 mmol) were addedsequentially at ambient temperature. Then excess diisopropylethylamine(1.2 mL, 6.80 mmol) was added dropwise. The reaction was stirred at roomtemperature for 18 hours and then concentrated under reduced pressure.The resulting residue was purified by reverse phase flash C18chromatography, elution gradient 20 to 75% acetonitrile in (watercontaining 0.2% NH₄OH), to afford6-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine(45 mg, 29%) as a solid. ¹H NMR (500 MHz, DMSO-d₆, 27° C.) 1.04 (3H, d),1.60-1.75 (2H, m), 2.51-2.70 (3H, m), 2.79-2.96 (3H, m), 3.03 (1H, dd),3.13 (1H, br dd), 3.41-3.53 (1H, m), 3.74 (2H, br s), 3.98 (1H, br d),4.45 (2H, dt), 4.86 (1H, s), 5.83 (1H, tt), 6.26 (1H, d), 6.74 (1H, d),6.81 (1H, dd), 6.96 (1H, d), 7.18 (1H, d), 7.76 (1H, d), 8.04 (1H, s),12.95 (1H, s). m/z: (ES+), [M+H]+ 459.

Procedures used to prepare the starting material tert-butyl3-((6-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-yl)amino)azetidine-1-carboxylateare described below.

Preparation of(1S,3R)-1-(5-bromopyridin-2-yl)-2-(2,2-difluoroethyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine

5-Bromopicolinaldehyde (2.74 g, 14.7 mmol) was added to a solution of(R)-3-(2-((2,2-difluoroethyl)amino)propyl)-2-methylaniline (1.6 g, 7.0mmol) in acetic acid (34.4 mL) and water (0.631 mL, 35.0 mmol). Thereaction was heated at 80° C. for 3 hours and then concentrated underreduced pressure. The resulting residue was dissolved in MeOH (40 mL)and sodium acetate (1.15 g, 14.0 mmol) and hydroxylamine hydrochloride(0.730 g, 10.5 mmol) were added. The reaction was stirred at roomtemperature for 3 hours and then concentrated under reduced pressure.The residue was dissolved in water, neutralized with saturated aqueousNaHCO₃, and then extracted with EtOAc. The combined organic layers wereconcentrated under reduced pressure, and the resulting residue waspurified by flash silica chromatography, elution gradient 0 to 90% EtOAcin hexanes. Product fractions were concentrated under reduced pressure,and the resulting residue was further purified by flash silicachromatography, elution gradient 0 to 90% EtOAc in hexanes, to afford(1S,3R)-1-(5-bromopyridin-2-yl)-2-(2,2-difluoroethyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(1.40 g, 49%) as a gum. ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 0.99 (3H, d),1.93 (3H, s), 2.38-2.47 (1H, m), 2.51-2.62 (1H, m), 2.72 (1H, dd),2.92-3.14 (1H, m), 3.23-3.29 (1H, m), 4.65 (2H, s), 4.79 (1H, s), 5.98(1H, tt), 6.39 (2H, s), 7.22 (1H, d), 7.91 (1H, dd), 8.55 (1H, d). m/z:ES+ [M+H]+ 396.

Preparation of(6S,8R)-6-(5-bromopyridin-2-yl)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

Acetic acid (541 mg, 9.01 mmol) was added to a stirred solution of(1S,3R)-1-(5-bromopyridin-2-yl)-2-(2,2-difluoroethyl)-3,5-dimethyl-1,2,3,4-tetrahydroisoquinolin-6-amine(714 mg, 1.80 mmol) in CHCl₃ (8 mL). The reaction was cooled to 0° C.,and a solution of isopentyl nitrite (422 mg, 3.60 mmol) in CHCl₃ (1 mL)was added dropwise. The reaction was stirred at 0° C. for 2 hours andthen quenched by the slow addition of a solution of NaHCO₃ (1.5 g, 18mmol) in water (20 mL). The phases were separated, and the organic layerwas concentrated under reduced pressure. The resulting residue waspurified by flash silica chromatography, elution gradient 5 to 35% EtOAcin hexanes, to afford(6S,8R)-6-(5-bromopyridin-2-yl)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(438 mg, 60%) as a light brown solid. ¹H NMR (300 MHz, CDCl₃, 27° C.)1.13 (3H, d), 2.68-2.85 (1H, m), 2.91 (1H, dd), 2.98-3.17 (1H, m), 3.34(1H, dd), 3.47-3.63 (1H, m), 5.05 (1H, s), 5.63 (1H, tt), 6.87 (1H, d),7.19 (1H, d), 7.28 (1H, d), 7.71 (1H, dd), 8.04 (1H, d), 8.57 (1H, dd).Indazole NH not observed. m/z: ES+ [M+H]+ 405.

Preparation of(6S,8R)-6-(5-bromopyridin-2-yl)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

3,4-Dihydro-2H-pyran (0.294 mL, 3.23 mmol) was added to a solution of(6S,8R)-6-(5-bromopyridin-2-yl)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(438 mg, 1.08 mmol) and para-toluenesulfonic acid monohydrate (21 mg,0.11 mmol) in DCM (4 mL). The reaction was heated at 100° C. for 6 hoursunder microwave conditions (300 W). The reaction was then allowed tocool and then washed with saturated aqueous NaHCO₃, dried over Na₂SO₄,filtered, and concentrated under reduced pressure. The resulting residuewas purified by flash silica chromatography, elution gradient 10 to 30%EtOAc in hexanes, to afford(6S,8R)-6-(5-bromopyridin-2-yl)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(441 mg, 83%) as a light brown gummy solid. m/z: ES+[M+H]+ 491.

Preparation of tert-butyl3-((64(6S,8R)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-yl)amino)azetidine-1-carboxylate

A vial was charged with a stirrer bar,(6S,8R)-6-(5-bromopyridin-2-yl)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.22 g, 0.44 mmol), tert-butyl 3-aminoazetidine-1-carboxylate (0.151 g,0.88 mmol), cesium carbonate (0.29 g, 0.88 mmol), and BrettPhos 3^(rd)Generation Precatalyst (0.040 g, 0.040 mmol). The vial was sealed,evacuated, and filled with nitrogen. 1,4-Dioxane (4 mL) was added, andthe vial was again evacuated and backfilled with nitrogen. The reactionwas stirred at 110° C. for 13 hours and then concentrated under reducedpressure. The resulting residue was purified by flash silicachromatography, elution gradient 10 to 30% EtOAc in hexanes, to affordtert-butyl3-((6-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-yl)amino)azetidine-1-carboxylate(0.20 g, 78%) as a beige solid. ¹H NMR (300 MHz, CD₃OD, 27° C.) 1.10(3H, dd), 1.43 (9H, s), 1.56-1.72 (2H, m), 1.72-1.87 (1H, m), 1.92-1.99(1H, m), 2.03-2.17 (1H, m), 2.38-2.53 (1H, m), 2.62-2.82 (1H, m),2.88-3.14 (2H, m), 3.33-3.44 (1H, m), 3.49-3.60 (1H, m), 3.67-3.83 (3H,m), 3.92-4.02 (1H, m), 4.17-4.33 (3H, m), 4.91 (1H, s), 5.34-5.57 (1H,m), 5.69-5.77 (1H, m), 6.79 (1H, d), 6.89 (1H, dd), 7.05 (1H, dd), 7.34(1H, d), 7.78-7.81 (1H, m), 8.07 (1H, s), aniline NH, not observed. m/z:ES+ [M+H]+ 583.

Example 26 Preparation of(6S,8R)-7-(2,2-difluoroethyl)-6-(5-O-(3-fluoropropyl)azetidin-3-yl)oxy)pyridin-2-yl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

HCl in dioxane (4N; 0.97 mL, 3.9 mmol) was added dropwise to a solutionof tert-butyl3-((6-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-yl)oxy)azetidine-1-carboxylate(0.227 g, 0.39 mmol) in MeOH (3 mL), and the reaction was stirred atroom temperature for 18 hours. The reaction was concentrated underreduced pressure to afford(6S,8R)-6-(5-(azetidin-3-yloxy)pyridin-2-yl)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolinehydrochloride (0.235 g; number of equivalents of HCl not determined) asa solid, which was used without further purification. m/z: (ES+), [M+H]+400.

1-Fluoro-3-iodopropane (0.041 mL, 0.34 mmol) was added to a stirredsolution of(6S,8R)-6-(5-(azetidin-3-yloxy)pyridin-2-yl)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolinehydrochloride (235 mg) in DMF (0.6 mL) at ambient temperature. Excessdiisopropylethylamine (1.36 mL, 7.80 mmol) was added dropwise. Thereaction was stirred for 18 hours at room temperature and thenconcentrated under reduced pressure. The resulting residue was purifiedby reverse phase flash C18 chromatography, elution gradient 20 to 75%acetonitrile in (water containing 0.2% NH₄OH). Product fractions werecombined and lyophilized to afford(6S,8R)-7-(2,2-difluoroethyl)-6-(5-((1-(3-fluoropropyl)azetidin-3-yl)oxy)pyridin-2-yl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(53 mg, 30%) as a solid. ¹H NMR (500 MHz, DMSO-d₆, 27° C.) 1.05 (3H, d),1.60-1.77 (2H, m), 2.54-2.70 (3H, m), 2.85 (1H, dd), 2.97-3.22 (4H, m),3.39-3.50 (1H, m), 3.87 (2H, br s), 4.45 (2H, dt), 4.80-4.92 (1H, m),5.00 (1H, s), 5.93 (1H, tt), 6.78 (1H, d), 7.19-7.26 (3H, m), 8.05 (1H,s), 8.06 (1H, d), 12.97 (1H, s). m/z: ES+ [M+H]+ 460.

Procedures used to prepare the starting material tert-butyl3-((6-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-yl)oxy)azetidine-1-carboxylateare described below.

Preparation of tert-butyl3-((64(6S,8R)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-yl)oxy)azetidine-1-carboxylate

Tert-butyl 3-hydroxyazetidine-1-carboxylate (227 mg, 1.31 mmol),(6S,8R)-6-(5-bromopyridin-2-yl)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(215 mg, 0.44 mmol), RockPhos 3rd Generation Precatalyst (37.1 mg, 0.04mmol), and cesium carbonate (285 mg, 0.88 mmol) were suspended intoluene (4 mL) in a sealed microwave vial. The reaction was heated at110° C. for 1 hour under microwave conditions (300 W). The reaction wasconcentrated under reduced pressure, and the resulting residue waspurified by flash silica chromatography, elution gradient 20 to 60%EtOAc in hexanes, to afford tert-butyl3-((6-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-3-(tetrahydro-2H-pyran-2-yl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-yl)oxy)azetidine-1-carboxylate(227 mg, 89%) as a solid. ¹H NMR (300 MHz, CD₃OD, 27° C.) 1.11 (3H, dd),1.41-1.43 (9H, m), 1.61-1.66 (1H, m), 1.66-1.86 (2H, m), 2.04-2.17 (1H,m), 2.36-2.54 (1H, m), 2.59-2.79 (1H, m), 2.89-3.02 (1H, m), 3.03-3.17(1H, m), 3.31-3.43 (1H, m), 3.48-3.60 (1H, m), 3.70 (2H, dd), 3.88-3.95(2H, m), 4.09-4.13 (1H, m), 4.27-4.39 (2H, m), 4.99-5.08 (2H, m), 5.64(1H, tt), 5.73 (1H, dt), 6.81 (1H, dd), 7.20 (1H, dd), 7.27 (1H, dd),7.36 (1H, d), 8.04-8.10 (2H, m). m/z: ES+ [M+H]+ 584.

Example 271-(3-Fluoropropyl)-N-(4-((6R,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-amine

Sodium tert-butoxide (2.118 g, 22.06 mmol) and BrettPhos G3 (0.166 g,0.18 mmol) were added to a degassed solution of1-(3-fluoropropyl)azetidin-3-amine (0.632 g, 4.78 mmol) and(6R,8R)-6-(4-bromophenyl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(1.56 g, 3.68 mmol) in 1,4-dioxane (18.4 ml) and the reaction was warmedto 90° C. and stirred overnight. After cooling, the reaction was dilutedwith EtOAc (20 mL) and water (20 mL), and the layers were separated. Theaqueous was extracted with EtOAc (20 mL), then the combined organicswere dried and evaporated. Purification was by HPLC (Waters CSH C18 OBDcolumn, 5μ silica, 30 mm diameter, 100 mm length), using decreasinglypolar mixtures of water (containing 0.1% NH₃ and MeCN as eluents) togive a gum. This was taken up in methanol (5 mL), then water (95 mL) wasadded and the mixture was slurried overnight at room temperature. Theresulting solid was collected by filtration, washed with 5% methanol inwater and dried in a vac oven at 50° C. overnight to give1-(3-fluoropropyl)-N-(4-((6R,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-amine(0.935 g, 54%) as a colourless solid. ¹H NMR (500 MHz, CDCl₃, 27° C.)1.13 (3H, d), 1.68-1.83 (2H, m), 2.59 (2H, t), 2.77 (1H, dd), 2.86 (2H,t), 2.89-2.99 (1H, m), 3.04 (1H, dd), 3.15-3.26 (1H, m), 3.37-3.48 (1H,m), 3.71-3.76 (2H, m), 3.93 (1H, d), 4.10 (1H, q), 4.43 (1H, t), 4.53(1H, t), 4.99 (1H, s), 6.41-6.46 (2H, m), 6.97 (1H, d), 7.00 (2H, d),7.26 (1H, s), 8.06 (1H, d), 10.10 (1H, s); m/z: ES+ [M+H]+ 476.

The(6R,8R)-6-(4-bromophenyl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineused as starting material was synthesised as follows:

(6R,8R)-6-(4-bromophenyl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

Trifluoroacetic acid (1.85 mL) was added to a solution of(R)-1-(1H-indazol-4-yl)-N-(2,2,2-trifluoroethyl)propan-2-amine (2.0 g,7.77 mmol) and 4-bromobenzaldehyde (7.19 g, 38.9 mmol) in toluene (37mL) and the resulting mixture was stirred at 90° C. for 30 hours. Thereaction was allowed to cool and partitioned between DCM (100 mL) andsaturated aqueous sodium bicarbonate (50 mL). The layers were separatedand the organic layer concentrated in vacuo. Purification was by silicagel column chromatography eluting with 0-50% ethyl acetate in heptane togive(6R,8R)-6-(4-bromophenyl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(1.560 g, 47%) as a foam, and as a 6:1 ratio of isomers. ¹H NMR (500MHz, CDCl₃, 27° C.) 1.14 (3H, d), 2.80 (1H, dd), 2.94 (1H, dd), 3.05(1H, dd), 3.25 (1H, dd), 3.33 (1H, ddd), 5.04 (1H, s), 6.95 (1H, d),7.08-7.12 (2H, m), 7.28-7.33 (1H, m), 7.37-7.40 (2H, m), 8.08 (1H, d),10.12 (1H, s); m/z: ES− [M−H]− 422.

Example 28 Preparation of1-(3-fluoropropyl)-N-(3-methoxy-44(6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-amine

BrettPhos 3rd Generation Precatalyst (10 mg, 0.01 mmol) and sodiumtert-butoxide (0.127 g, 1.32 mmol) were added in one portion to adegassed solution of 1-(3-fluoropropyl)azetidin-3-amine (0.033 g, 0.25mmol) and(6S,8R)-6-(4-bromo-2-methoxyphenyl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(0.10 g, 0.22 mmol) in 1,4-dioxane (1.10 mL). The orange mixture wasthen immersed in an oil bath that had been preheated to 50° C. After 5minutes, the reaction was cooled to room temperature. In a separateflask, sodium tert-butoxide (1.81 g, 18.8 mmol) and BrettPhos 3rdGeneration Precatalyst (0.17 g, 0.19 mmol) were added in one portion toa degassed solution of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(1.71 g, 3.76 mmol) and 1-(3-fluoropropyl)azetidin-3-amine (0.597 g,4.52 mmol) in 1,4-dioxane (18.8 mL). The light orange mixture wasimmersed in an oil bath that had been preheated to 50° C. After 5minutes, the reaction was cooled to room temperature. Once cooled, bothreactions were combined, diluted with ethyl acetate and washedsequentially with water (×2) and saturated aqueous sodium chloride. Theorganic layer was dried over sodium sulfate, filtered and concentratedunder reduced pressure. The resulting orange oil was purified by flashsilica chromatography, elution gradient 0 to 10% methanol in DCM, toafford1-(3-fluoropropyl)-N-(3-methoxy-4-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-amine(1.84 g, 92%) as a solid and an ˜84:16 trans:cis mixture based on UVHPLC profile. This material was resolved using preparative SFC (column:Chiralpak AD, 21.2×250 mm, 5 μm; 75 mL/min), eluting with 20% (methanolcontaining 0.2% NH₄OH) in CO₂, to afford1-(3-fluoropropyl)-N-(3-methoxy-4-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-amine(1.01 g) as a light orange solid and a second eluting peak. Thismaterial was further purified by flash silica chromatography, elutiongradient 0 to 30% methanol in ethyl acetate, to afford1-(3-fluoropropyl)-N-(3-methoxy-4-46S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-amine(0.954 g) as a white solid. This material was purified a final time bypreparative HPLC (column: Xbridge C18, 30×100 mm, 5 μm, 40 mL/min),eluting with 40 to 70% acetonotrile in (water containing 0.2% NH₄OH).Product fractions were combined, washed with ethyl acetate (×3), and thecombined organic layers were washed with saturated aqueous sodiumchloride, dried over sodium sulfate, filtered, and concentrated underreduced pressure to afford1-(3-fluoropropyl)-N-(3-methoxy-4-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-amine(551 mg, 27%) as an off-white foam solid. ¹H NMR (500 MHz, DMSO-d₆, 27°C.) 1.04 (3H, d), 1.63 (2H, dtt), 2.44 (2H, t), 2.70 (2H, dd), 2.81 (1H,dd), 2.84-2.94 (1H, m), 3.09 (1H, dd), 3.34 (1H, br s), 3.44 (1H, dqd),3.61 (2H, dd), 3.77 (3H, s), 3.90 (1H, m), 4.43 (2H, dt), 5.28 (1H, s),5.87 (1H, dd), 6.02 (1H, d), 6.16 (1H, d), 6.32 (1H, d), 6.65 (1H, d),7.18 (1H, d), 8.02 (1H, s), 12.95 (1H, s). m/z: ES+ [M+H]+ 506.

Procedures used to prepare the starting material(6S,8R)-6-(4-bromo-2-methoxyphenyl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineare described below.

Preparation of(R)—N-(1-(3-amino-2-methylphenyl)propan-2-0)-2,2,2-trifluoroacetamide

Ethyl 2,2,2-trifluoroacetate (3.75 ml, 31.5 mmol) was added to a darkamber-red solution of (R)-3-(2-aminopropyl)-2-methylaniline (5.17 g,31.5 mmol) and triethylamine (4.83 mL, 34.6 mmol) in MeOH (70.1 mL).After 15 min, the reaction was concentrated to a dark amber oil. The oilwas dissolved in ethyl acetate, washed with water, and the aqueous layerextracted with ethyl acetate (×2). The combined organic layers weredried over sodium sulfate, filtered and concentrated under reducedpressure. The resulting oil was purified by flash silica chromatography,elution gradient 0 to 40% ethyl acetate in hexanes, to afford(R)—N-(1-(3-amino-2-methylphenyl)propan-2-yl)-2,2,2-trifluoroacetamide(6.47 g, 79%) as a yellow-orange oil that crystallized to a light orangesolid on standing. ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 1.11 (3H, d), 2.01(3H, s), 2.63 (1H, dd), 2.78 (1H, dd), 3.93-4.05 (1H, m), 4.71 (2H, s),6.36 (1H, dd), 6.49 (1H, dd), 6.78 (1H, t), 9.25 (1H, br d). m/z: ES+[M+H]+ 261.

Preparation of(R)-2-methyl-3-(2-((2,2,2-trifluoroethyl)amino)propyl)aniline

Borane tetrahydrofuran complex in THF (1 M; 149 mL, 149 mmol) was addedvia syringe (6×25 mL) to a solution of(R)—N-(1-(3-amino-2-methylphenyl)propan-2-yl)-2,2,2-trifluoroacetamide(6.47 g, 24.9 mmol) in tetrahydrofuran (81 mL) at 0° C. The ice bath wasremoved, and, upon warming, gas evolution was observed. Once no furthergas evolution was visible (˜20 minutes), the reaction was warmed to 65°C. After 4 hours, the reaction was cooled and maintained at roomtemperature for 18 hours. The reaction was then cooled to 0° C. andquenched by dropwise addition of methanol (22 mL). The reaction waswarmed to room temperature, and, after gas evolution ceased, thereaction was warmed to 65° C. After 14 hours, the reaction was thencooled to room temperature and stirred under these conditions for 3.5days before being concentrated under reduced pressure. The resulting oilwas purified by flash silica chromatography, elution gradient 0 to 70%ethyl acetate in hexanes, to afford(R)-2-methyl-3-(2-((2,2,2-trifluoroethyl)amino)propyl)aniline (5.62 g,92%) as a light yellow oil. ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 0.90 (3H,d), 1.98 (3H, s), 2.09-2.20 (1H, m), 2.25-2.40 (1H, m), 2.72-2.84 (2H,m), 3.17-3.29 (2H, m), 4.69 (2H, s), 6.36 (1H, dd), 6.49 (1H, dd), 6.78(1H, t). m/z: ES+ [M+H]+ 247

Preparation of(1S,3R)-1-(4-Bromo-2-methoxyphenyl)-3,5-dimethyl-2-(2,2,2-trifluoroethyl)-1,2,3,4-tetrahydroisoquinolin-6-amine

4-Bromo-2-methoxybenzaldehyde (1.83 g, 8.53 mmol) and(R)-2-methyl-3-(2-((2,2,2-trifluoroethyl)amino)propyl)aniline (2.00 g,8.12 mmol) were added to a solution of water (0.7 mL, 41 mmol) andacetic acid (40 mL). The resulting light yellow solution was immersed inan oil bath that had been preheated to 65° C. and was maintained underthese conditions for 18 hours. The resulting dark amber solution wasthen concentrated under reduced pressure (water bath: 55° C.). Theresulting residue was diluted with ethyl acetate and washed withsaturated aqueous sodium hydrogencarbonate until the aqueous layer wasconfirmed to be pH=8 using a pH strip. The organic layer was dried oversodium sulfate, filtered, and concentrated under reduced pressure. Theresulting residue was dissolved in methanol (20 mL) and hydroxylaminehydrochloride (0.152 g, 2.19 mmol) and excess potassium carbonate wereadded. After 5 minutes, the mixture was diluted with ethyl acetate,filtered, and the filtrate was concentrated under reduced pressure. Theresulting residue was purified by flash silica chromatgraphy, elutiongradient 0 to 50% ethyl acetate in hexanes, to afford(1S,3R)-1-(4-bromo-2-methoxyphenyl)-3,5-dimethyl-2-(2,2,2-trifluoroethyl)-1,2,3,4-tetrahydroisoquinolin-6-amineas a dark amber oil in an 83:17 trans:cis ratio based on NMRintegration. The oil was reconcentrated from DCM and dried under vacuumto a light amber foam solid (2.55 g, 71%). A small amount of thismaterial (150 mg) was resolved using preparative SFC (column: (S,S)Whelk-01, 21.2×250 mm, 5 μm; 75 mL/min), eluting with 15% (methanolcontaining 0.2% NH₄OH) in CO₂, to afford slower eluting(1S,3R)-1-(4-bromo-2-methoxyphenyl)-3,5-dimethyl-2-(2,2,2-trifluoroethyl)-1,2,3,4-tetrahydroisoquinolin-6-amine(101 mg) and faster eluting(1R,3R)-1-(4-bromo-2-methoxyphenyl)-3,5-dimethyl-2-(2,2,2-trifluoroethyl)-1,2,3,4-tetrahydroisoquinolin-6-amine(18 mg) as faint yellow foam solids.

(1S,3R): ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 1.00 (3H, d), 1.95 (3H, s),2.43 (1H, dd), 2.64-2.85 (2H, m), 3.19-3.43 (2H, m), 3.86 (3H, s), 4.67(2H, s), 5.18 (1H, s), 6.29 (1H, d), 6.41 (1H, d), 6.60 (1H, d), 6.96(1H, dd), 7.18 (1H, d). m/z: ES+ [M+H]+ 443.

(1R,3R): ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 1.19 (3H, d), 1.94 (3H, s),2.53-2.60 (1H, m), 2.78 (1H, br dd), 2.98-3.12 (1H, br m), 3.16-3.36(2H, m), 3.84 (3H, s), 4.61 (2H, s), 5.31 (1H, s), 6.20 (1H, d), 6.33(1H, d), 7.03-7.13 (2H, m), 7.19 (1H, d). m/z: ES+ [M+H]+443.

Preparation of(6S,8R)-6-(4-bromo-2-methoxyphenyl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

A solution of sodium nitrite (0.413 g, 5.98 mmol) in water (3.83 mL) wasadded dropwise over 5 minutes to a stirred solution of(1S,3R)-1-(4-bromo-2-methoxyphenyl)-3,5-dimethyl-2-(2,2,2-trifluoroethyl)-1,2,3,4-tetrahydroisoquinolin-6-amine(2.55 g, 5.75 mmol; 83:17 trans:cis) in propionic acid (19.2 mL)maintained at −15° C. using a cooling bath consisting ice, solid sodiumchloride, and saturated aqueous sodium chloride. After 20 minutes thereaction was diluted with toluene (100 mL) that had been precooled to−70° C. The resulting light yellow mixture was stirred vigorously, and,after 5 minutes, the cooling bath was removed. Upon reaching roomtemperature, the red reaction mixture was maintained under theseconditions for 1.5 hours and then washed with water (×2). The combinedaqueous layers were extracted with ethyl acetate. The combined organiclayers were dried over sodium sulfate, filtered, and concentrated underreduced pressure to approximately 20% of the starting volume. Thismixture was diluted with EtOAc (20 mL) and washed with saturated aqueoussodium bicarbonate. Solid potassium carbonate was then added until gasevolution ceased and the mixture tested as being basic using a pH strip.The layers were separated and the organic layer was dried over sodiumsulfate, filtered and concentrated under reduced pressure. The resultingoil was purified by flash silica chromatography, elution gradient 0 to70% ethyl acetate in hexanes, to afford(6S,8R)-6-(4-bromo-2-methoxyphenyl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(1.96 g, 75%) as a light orange foam solid and a ˜9:1 trans:cis mixturebased on NMR integration. ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 1.06 (3H,d), 2.80-2.98 (2H, m), 3.15 (1H, br dd), 3.33-3.53 (2H, m), 3.90 (3H,s), 5.40 (1H, s), 6.63 (1H, d), 6.67 (1H, d), 6.96 (1H, dd), 7.20-7.27(2H, m), 8.08 (1H, s), 13.00 (1H, s). m/z: ES+ [M+H]+ 454.

Example 29 Preparation of2-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine

Sodium tert-butoxide (1.08 g, 11.3 mmol) and BrettPhos 3rd GenerationPrecatalyst (0.16 g, 0.18 mmol) were added to a degassed solution of(6S,8R)-6-(5-bromo-6-fluoropyridin-2-yl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(2.00 g, 4.51 mmol) and 1-(3-fluoropropyl)azetidin-3-amine (0.90 g, 5.41mmol) in 1,4-dioxane (22.6 mL) at room temperature. The red-orangemixture was immersed in an oil bath preheated to 44° C. After 22minutes, the orange mixture was removed from the heat and poured intoethyl acetate and saturated aqueous sodium chloride and the layers wereseparated. The aqueous layer was extracted with ethyl acetate, and thecombined organic layers were washed with saturated aqueous sodiumchloride, dried over sodium sulfate, filtered and concentrated underreduced pressure. The resulting residue was purified by flash silicachromatography, elution gradient 0 to 30% methanol in ethyl acetate, toafford2-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine(1.58 g) as a light yellow foam solid and an approximate 84:16 trans:cismixture based on NMR integration. This material was resolved bypreparative SFC (column: Lux Cellulose-4, 21.2×250 mm, 5 μm; 70 mL/min),eluting with 35% (methanol containing 0.2% NH₄OH) in CO₂, to afford alight orange foam solid. This solid was repurified by flash silicachormatography, elution gradient 0 to 30% methanol in ethyl acetate.Product fractions were concentrated under reduced pressure. Theresulting residue was taken up in acetonitrile, filtered, andconcentrated under reduced pressure to afford2-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine(1.03 g, 46%) as a light yellow foam solid. ¹H NMR (600 MHz, DMSO-d₆,27° C.) 1.10 (3H, d), 1.64 (2H, dtt), 2.48 (2H, t), 2.83 (1H, dd), 2.86(2H, br d), 2.97 (1H, br dq), 3.02 (1H, dd), 3.45 (1H, dqd), 3.54 (1H,dq), 3.63 (2H, br s), 3.96 (1H, dquin), 4.44 (2H, dt), 4.91 (1H, s),6.13 (1H, br d), 6.87 (1H, d), 6.92-7.01 (2H, m), 7.25 (1H, d), 8.06(1H, s), 13.00 (1H, s). m/z: ES+ [M+H]+ 495.

Procedures used to prepare the starting material(6S,8R)-6-(5-bromo-6-fluoropyridin-2-yl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineare described below.

Preparation of tert-butyl (R)-(1-(1H-indazol-4-yl)propan-2-yl)carbamate

n-Butyllithium in hexane (2.5 M; 96 mL, 241 mmol) was added to asolution of 4-bromo-1H-indazole (24.8 g, 126 mmol) in THF (200 mL) at−78° C. over 20 minutes, and the mixture was stirred at −78° C. for 7hours. Tert-butyl (R)-4-methyl-1,2,3-oxathiazolidine-3-carboxylate2,2-dioxide (26 g, 110 mmol) was added, and the resultant mixture wasstirred at −78° C. for 15 minutes. The cooling bath was removed, and themixture was stirred under these conditions for 18 hours. Aqueuous citricacid (1N; 130 mL) was added and stirring was continued for 30 minutes.The mixture was extracted with hexanes, and the organic layer was washedwith saturated aqueous sodium carbonate, dried over sodium sulfate,filtered and concentrated under reduced pressure. The resulting residuewas purified by flash silica chromatography, eluting with 0 to 60% EtOAcin hexanes, to give tert-butyl(R)-(1-(1H-indazol-4-yl)propan-2-yl)carbamate (17.3 g, 57%) as a whitesolid. ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 1.02 (3H, br d), 1.34 (9H, s),2.82 (1H, br dd), 3.09 (1H, br dd), 3.82 (1H, dt), 6.82 (1H, br d), 6.88(1H, d), 7.24 (1H, dd), 7.35 (1H, br d), 8.18 (1H, s), 12.97 (1H, s).m/z: ES+ [M+1]+276.

Preparation of (R)-1-(1H-indazol-4-yl)propan-2-amine dihydrochloride

HCl dioxane (4 M, 100 mL, 400 mmol) was added to a suspension oftert-butyl (R)-(1-(1H-indazol-4-yl)propan-2-yl)carbamate (17.3 g, 62.7mmol) in DCM (200 mL) at room temperature over 10 minutes. The resultingslurry was stirred overnight and then concentrated under reducedpressure to afford (R)-1-(1H-indazol-4-yl)propan-2-amine (15.9 g, 102%)as white solid. ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 1.14 (3H, d), 2.97(1H, dd), 3.40 (1H, dd), 3.45-3.62 (1H, m), 6.96 (1H, d), 7.29 (1H, dd),7.45 (1H, d), 7.97-8.27 (3H, br s), 8.29 (1H, d). m/z: ES+ [M+H]+ 176.

Preparation of(R)-1-(1H-indazol-4-yl)-N-(2,2,2-trifluoroethyl)propan-2-amine

Potassium carbonate (4.75 g, 34.3 mmol) was added to a stirredsuspension of (R)-1-(1H-indazol-4-yl)propan-2-amine dihydrochloride salt(2.13 g, 8.58 mmol) in acetonitrile (25 mL) then 2,2,2-trifluoroethyltrifluoromethanesulfonate (2.191 g, 9.44 mmol) in DCM (12.6 mL) wasadded dropwise. The mixture was stirred at room temperature for 1.5days. Then additional 2,2,2-trifluoroethyl trifluoromethanesulfonate(398 mg) in DCM (0.3 mL) was added. The reaction was warmed to 60° C.and after 3 hours, the reaction was cooled to room temperature and afurther portion of 2,2,2-trifluoroethyl trifluoromethanesulfonate (398mg) was added as a solution in DCM (0.3 mL). After 18 hours, thereaction was concentrated under reduced pressure to reduced volume andthen diluted with DCM. The mixture was washed with water, and theorganic layer was dried over sodium sulfate, filtered and concentratedunder reduced pressure. The resulting residue was purified by flashsilica chromatography, elution gradient 0 to 60% ethyl acetate inhexanes, to afford(R)-1-(1H-indazol-4-yl)-N-(2,2,2-trifluoroethyl)propan-2-amine (2.06 g,93%) as a gum. m/z: ES+[M+H]+ 257.

Preparation of(6S,8R)-6-(5-bromo-6-fluoropyridin-2-yl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

Trifluoroacetic acid (1.7 mL) was added to a solution of(R)-1-(1H-indazol-4-yl)-N-(2,2,2-trifluoroethyl)propan-2-amine (1.83 g,7.11 mmol) and 5-bromo-6-fluoropicolinaldehyde (1.45 g, 7.11 mmol) intoluene (33.8 mL). The reaction was heated at 90° C. for 24 hours andthen concentrated to reduced volume. The mixture was then diluted withdichloromethane and basified with saturated aqueous sodiumhydrogencarbonate. The organic layer was dried over sodium sulfate,filtered and concentrated under reduced pressure. The resulting residuewas purified by flash silica chromatography, elution gradient 0 to 50%ethyl acetate in hexanes, to afford(6S,8R)-6-(5-bromo-6-fluoropyridin-2-yl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(2.01 g, 64%) as a white solid. ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 1.11(3H, d), 2.89 (1H, br dd), 2.94-3.09 (2H, m), 3.31-3.42 (1H, m),3.52-3.71 (1H, m), 5.07 (1H, s), 6.96 (1H, d), 7.24-7.36 (2H, m), 8.06(1H, d), 8.23 (1H, dd), 13.02 (1H, s). m/z: ES+[M+H]+ 443.

Example 305-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine

1-(3-Fluoropropyl)azetidin-3-amine (435 mg, 3.29 mmol),(6S,8R)-6-(5-bromo-3-fluoropyridin-2-yl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(730 mg, 1.65 mmol) and sodium tert-butoxide (950 mg, 9.88 mmol) weresuspended in 1,4-dioxane (18.3 mL). The mixture was degassed andBrettphos 3G precatalyst (149 mg, 0.16 mmol) was added. The reaction washeated to 80° C. for 1 hour. The reaction mixture was cooled to roomtemperature and diluted with ethyl acetate (10 mL) and washed with water(10 mL). The organic layer was dried over magnesium sulfate, filteredand concentrated under vacuum. The crude product was purified bypreparative LCMS (Waters XSelect CSH C18 column, 5μ silica, 50 mmdiameter, 100 mm length), using decreasingly polar mixtures of water(containing 1% NH₃) and MeCN as eluents. The sample was dissolved inMeOH and separated by SFC using the following chromatographc conditions:Column: Phenomonex Lux C1, 30×250 mm, 5 micron, Mobile phase: 30%MeOH+0.1% NH₃/70% scCO₂ to afford5-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine(396 mg, 49%) as a yellow solid. ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.15(3H, d), 1.69-1.81 (2H, m), 2.60 (2H, t), 2.77 (1H, dd), 2.92-3.03 (3H,m), 3.14-3.31 (2H, m), 3.65-3.79 (3H, m), 4.04 (1H, q), 4.43 (2H, t),4.53 (1H, t), 5.35 (1H, s), 6.54 (1H, dd), 6.76 (1H, d), 7.00 (1H, d),7.66 (1H, d), 7.93 (1H, d), 11.06 (1H, s); m/z: ES+ [M+H]+ 495.

The(6S,8R)-6-(5-bromo-3-fluoropyridin-2-yl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineused as starting material was synthesised as follows:

(6S,8R)-6-(5-bromo-3-fluoropyridin-2-yl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

Trifluoroacetic acid (2.13 mL) was added to a solution of(R)-1-(1H-indazol-4-yl)-N-(2,2,2-trifluoroethyl)propan-2-amine (1.15 g,4.47 mmol) and 5-bromo-3-fluoropicolinaldehyde (912 mg, 4.47 mmol) intoluene (42.6 mL) and the resulting mixture was stirred at 100° C. for30 minutes. The reaction was evaporated and the residue partitionedbetween DCM (20 mL) and 2M NaOH (20 mL). The layers were separated andthe organic phase was concentrated under reduced pressure. The crudeproduct was purified by flash silica chromatography, elution gradient 0to 40% EtOAc in heptane. Product containing fractions were evaporated todryness to afford(6S,8R)-6-(5-bromo-3-fluoropyridin-2-yl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(1.15 g, 58%) as a yellow solid. ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.16(3H, d), 2.86 (1H, dd), 3.01 (1H, dq), 3.19-3.35 (2H, m), 3.68-3.78 (1H,m), 5.42 (1H, s), 6.80 (1H, d), 7.20 (1H, d), 7.59 (1H, dd), 8.05 (1H,d), 8.27-8.5 (1H, m). m/z: ES+ [M+H]+ 443.

Example 312,2-Difluoro-3-((6S,8R)-6-(3-fluoro-5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,34]isoquinolin-7-yl)propan-1-ol

Tetrabutylammonium fluoride (1.0 M in THF, 0.65 mL, 0.65 mmol) was addedto a solution of6-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-5-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine(320 mg, 0.43 mmol) in THF (2.66 mL) at room temperature and stirred for64 hours. The reaction mixture was evaporated and then dissolved in DMSOand the crude product was purified by flash reverse phase (Puriflash,220 g, C18, 30μ column), using decreasingly polar mixtures of water(containing 1% NH₃) and MeCN as eluents. Fractions containing thedesired compound were evaporated to dryness to afford a crude product(143 mg). The crude product was purified by flash silica chromatography,elution gradient 0 to 20% MeOH in EtOAc. Product containing fractionswere evaporated to dryness to afford2,2-difluoro-3-((6S,8R)-6-(3-fluoro-5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol(118 mg, 54%) as a mixture of isomers. The material was repurified bypreparative HPLC (Waters XSelect CSH C18 column, 5μ silica, 30 mmdiameter, 100 mm length), using decreasingly polar mixtures of water(containing 0.1% formic acid) and MeCN as eluents. Fractions containingthe desired compound were combined and purified by ion exchangechromatography, using an SCX-2 column. The desired product was elutedfrom the column using 1M NH₃/MeOH and product containing fractions wereevaporated to dryness to afford2,2-difluoro-3-((6S,8R)-6-(3-fluoro-5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol(62.1 mg, 29%) as a colourless solid. ¹H NMR (500 MHz, DMSO, 27° C.)1.03 (3H, dd), 1.64 (2H, dq), 2.44 (2H, t), 2.60-2.66 (1H, m), 2.79 (3H,dd), 2.97 (1H, dd), 3.04-3.15 (1H, m), 3.50-3.73 (5H, m), 3.89-3.97 (1H,m), 4.39 (1H, t), 4.48 (1H, t), 5.21 (1H, s), 5.26 (1H, t), 6.58 (1H,d), 6.65-6.71 (2H, m), 7.19 (1H, d), 7.55 (1H, dd), 8.03 (1H, s), 12.94(1H, s); m/z: ES+ [M+H]+ 507.

The6-((6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-5-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amineused as starting material was synthesised as follows:

(R)—N-(1-(1H-indazol-4-yl)propan-2-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropan-1-amine

3-((Tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyltrifluoromethanesulfonate (2.59 g, 5.36 mmol) was added to a solution of(R)-1-(1H-indazol-4-yl)propan-2-amine (0.94 g, 5.36 mmol) and DIPEA(1.39 ml, 8.05 mmol) in 1,4-dioxane (38.9 mL) and the reaction wasstirred at 50° C. for 18 hours. The reaction mixture was evaporated thenthe residue was diluted with EtOAc and washed with water and the aqueouslayer was further extracted with EtOAc. The combined organic layers werewashed with saturated brine, dried over magnesium sulfate, filtered andevaporated to dryness. The crude product was purified by flash silicachromatography, elution gradient 0 to 50% EtOAc in heptane. Purefractions were evaporated to dryness to afford(R)—N-(1-(1H-indazol-4-yl)propan-2-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropan-1-amine(1.515 g, 52%) as a colourless gum. ¹H NMR (500 MHz, DMSO, 27° C.) 0.92(3H, d), 0.97 (9H, s), 1.78-1.86 (1H, m), 2.73 (1H, dd), 2.98-3.14 (4H,m), 3.83 (2H, td), 6.85 (1H, d), 7.20 (1H, dd), 7.34 (1H, d), 7.41-7.50(6H, m), 7.58-7.64 (4H, m), 8.08 (1H, s), 12.98 (1H, s); m/z: ES+ [M+H]+508.

(6S,8R)-6-(5-bromo-3-fluoropyridin-2-yl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

Trifluoroacetic acid (319 μL) was added to a solution of(R)—N-(1-(1H-indazol-4-yl)propan-2-yl)-3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropan-1-amine(681 mg, 1.34 mmol) and 5-bromo-3-fluoropicolinaldehyde (287 mg, 1.41mmol) in toluene (6.39 mL) and the resulting mixture was stirred at 110°C. for 1 hour. The reaction was allowed to cool to room temperature,evaporated to dryness and dissolved in DMSO. The crude product waspurified byfFlash reverse phase chromatography (100 g Redisep Rf C18column), using decreasingly polar mixtures of water (containing 0.1%formic acid) and MeCN (60-100%) as eluents. Fractions containing thedesired compound were combined and was isolated by ion exchangechromatography, using an SCX-2 column. The desired product was elutedfrom the column using 1M NH₃/MeOH and pure fractions were evaporated todryness to afford(6S,8R)-6-(5-bromo-3-fluoropyridin-2-yl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(763 mg, 82%) as an off-white solid. ¹H NMR (500 MHz, DMSO, 27° C.) 0.99(9H, s), 1.04 (3H, d), 2.74-2.89 (2H, m), 2.97-3.04 (1H, m), 3.31 (1H,s), 3.54-3.62 (1H, m), 3.78 (1H, q), 3.92-4.02 (1H, m), 5.36 (1H, s),6.72 (1H, d), 7.23 (1H, d), 7.41-7.49 (6H, m), 7.57-7.61 (4H, m), 8.04(1H, dd), 8.09 (1H, s), 8.38 (1H, d), 13.01 (1H, s); m/z: ES+ [M+H]+693.

64(6S,8R)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-5-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine

1-(3-Fluoropropyl)azetidin-3-amine (114 mg, 0.86 mmol),(6S,8R)-6-(5-bromo-3-fluoropyridin-2-yl)-7-(3-((tert-butyldiphenylsilyl)oxy)-2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(300 mg, 0.43 mmol) and sodium tert-butoxide (249 mg, 2.59 mmol) weresuspended in degassed 1,4-dioxane (4.81 mL). Brettphos 3G precatalyst(39.2 mg, 0.04 mmol) was added and the mixture was evacuated and purgedwith nitrogen (×2) and the reaction was then heated to 80° C. for 45minutes. The reaction mixture was allowed to cool to room temperatureand diluted with EtOAc and washed with water. The aqueous layer wasfurther extracted with EtOAc and the combined organic layers were washedwith saturated brine, dried over magnesium sulfate, filtered andevaporated to dryness and used directly without further purification.m/z: ES+ [M+H]+ 745.

Example 326-((6S,8R)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,9,8-tetrahydro-3H-pyrazolo[4,3-f]isoquin-6-yl)-N-(3-fluoropropyl)azetidi-3-yl)pyridin-3-amine

Sodium tert-butoxide (116 mg, 1.20 mmol) and BrettPhos G3 (9.09 mg,10.04 μmol) were added to a degassed solution of(6S,8R)-6-(5-bromopyridin-2-yl)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(89 mg, 0.20 mmol) and 1-(3-fluoropropyl)azetidin-3-amine (39.8 mg, 0.30mmol) in 1,4-dioxane (1.00 mL) and reaction was warmed to 90° C. andstirred for 6 hours. After cooling, the reaction was diluted with EtOAcand water, and the layers were separated. The aqueous layer wasextracted with EtOAc, then the combined organics were dried over sodiumsulfate and evaporated. Purification was by HPLC (Waters CSH C18 OBDcolumn, 5μ silica, 30 mm diameter, 100 mm length), using decreasinglypolar mixtures of water (containing 0.1% NH₃ and MeCN as eluents) togive the product (42.0 mg, 42%) as a film. ¹H NMR (500 MHz, CDCl₃, 27°C.) 1.13 (3H, d), 1.69-1.84 (2H, m), 2.61 (2H, td), 2.76 (1H, dd), 2.94(3H, ddd), 3.16-3.28 (2H, m), 3.45-3.54 (1H, m), 3.67-3.76 (2H, m), 4.09(1H, dt), 4.24 (1H, d), 4.43 (1H, td), 4.53 (1H, td), 5.02 (1H, s), 6.76(1H, dd), 6.81 (1H, d), 6.89 (1H, dd), 7.39 is (1H, d), 7.81 (1H, d),11.10 (1H, s); m/z: ES+ [M+H]+ 495.

The(6S,8R)-6-(5-bromopyridin-2-yl)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineused as starting material was synthesised as follows:

4-Bromo-3-fluoro-1H-indazole

Selectfluor (49.40 g, 139.6 mmol) was added to a solution of4-bromo-1H-indazole (25.0 g, 127 mmol) in DMF (254 mL) and the reactionwas heated to 70° C. overnight. After cooling, the reaction mixture waspoured onto water. The precipitated solid was filtered and dried, thenthe crude product was purified by flash silica chromatography, elutiongradient 0 to 30% EtOAc in heptane. Product containing fractions wereevaporated to dryness to afford 4-bromo-3-fluoro-1H-indazole (4.20 g,15%) as a pale yellow solid. ¹H NMR (500 MHz, CDCl₃, 27° C.) 7.24-7.28(1H, m), 7.32-7.36 (2H, m), 9.20 (1H, s); m/z: ES− [M−H]− 213.

Tert-butyl (R)-(1-(3-fluoro-1H-indazol-4-yl)propan-2-yl)carbamate

n-Butyl lithium (1.6M, 51.9 ml, 83.01 mmol) was added to4-bromo-3-fluoro-1H-indazole (8.50 g, 39.5 mmol) in THF (124 mL) at −78°C. and the reaction was stirred for 15 minutes, warmed to −50° C. for 15minutes, then cooled back to −78° C. Tert-butyl(R)-4-methyl-1,2,3-oxathiazolidine-3-carboxylate 2,2-dioxide (10.32 g,43.48 mmol) was added in THF (20 mL) and the reaction was stirred for 15minutes before being allowed to warm to −10° C. over 30 minutes. 1Ncitric acid (200 mL) was added and the mixture was stirred for 15minutes, before being extracted with EtOAc (×2). The combined organicswere dried over magnesium sulfate and evaporated. The crude product waspurified by flash silica chromatography, elution gradient 0 to 40% EtOAcin heptane. Product containing fractions were evaporated to dryness toafford tert-butyl (R)-(1-(3-fluoro-1H-indazol-4-yl)propan-2-yl)carbamate(5.93 g, 51%) as a colourless solid. ¹H NMR (500 MHz, CDCl₃, 2 7° C.)1.17 (3H, d), 1.35 (9H, s), 3.09 (2H, d), 4.00-4.09 (1H, m), 4.41-4.49(1H, m), 6.97 (1H, d), 7.22 (1H, dd), 7.33 (1H, dd), 9.36 (1H, s); m/z:ES− [M−H]− 292.

(R)-1-(3-Fluoro-1H-indazol-4-yl)propan-2-amine

4N Hydrochloric acid in dioxane (23.86 ml, 95.45 mmol) was added totert-butyl (R)-(1-(3-fluoro-1H-indazol-4-yl)propan-2-yl)carbamate (5.60g, 19.1 mmol) in MeOH (23.9 mL) and the reaction was stirred at roomtemperature for 2 hours. The crude mixture was concentrated, thensuspended in EtOAc (100 mL) and washed with saturated aqueous sodiumbicarbonate (50 mL). The aqueous layer was extracted with EtOAc (×5),then the combined organics were dried over sodium sulfate and evaporatedto afford (R)-1-(3-fluoro-1H-indazol-4-yl)propan-2-amine (3.10 g, 84%)as a yellow oil, which solidified on standing.

¹H NMR (500 MHz, CDCl₃, 27° C.) 1.19 (3H, d), 2.86 (1H, dd), 3.09 (1H,dd), 3.33 (1H, dddd), 6.96 (1H, d), 7.22 (1H, dd), 7.33 (1H, dd), 9.93(1H, s); m/z: ES+ [M+H]+ 194.

(R)-1-(3-Fluoro-1H-indazol-4-yl)-N-(2,2,2-trifluoroethyl)propan-2-amine

2,2,2-Trifluoroethyl trifluoromethanesulfonate 0.1M solution in DCM(25.9 mL, 2.59 mmol) was added to(R)-1-(3-fluoro-1H-indazol-4-yl)propan-2-amine (0.4 g, 2.07 mmol) andDIPEA (0.541 mL, 3.11 mmol) in 1,4-dioxane (20 mL) and the resultingmixture was stirred at 75° C. overnight. The reaction was concentratedin vacuo and partitioned between ethyl acetate (25 mL) and saturatedaqueous sodium bicarbonate (25 mL). The layers were separated and theaqueous layer extracted with ethyl acetate (25 mL). The combinedorganics were washed with saturated aqueous sodium chloride (25 mL),dried over sodium sulphate, filtered and concentrated in vacuo. Theresulting gum was taken up in methanol and applied to a pre-wetted(methanol) SCX-2 cartridge. The cartridge was washed with methanol andeluted with 1M ammonia in methanol. The eluent was concentrated in vacuto give(R)-1-(3-fluoro-1H-indazol-4-yl)-N-(2,2,2-trifluoroethyl)propan-2-amine(0.455 g, 80%) as a brown gum. ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.11 (3H,d), 2.92 (1H, dd), 3.07-3.22 (4H, m), 6.96 (1H, d), 7.25 (1H, dd), 7.34(1H, dd), 9.50 (1H, s); m/z: ES+ [M+H]+276.

(6S,8R)-6-(5-bromopyridin-2-yl)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

Trifluroacetic acid (87 μL) was added to a solution of(R)-1-(3-fluoro-1H-indazol-4-yl)-N-(2,2,2-trifluoroethyl)propan-2-amine(100 mg, 0.36 mmol) and 5-bromopicolinaldehyde (67.6 mg, 0.36 mmol) intoluene (0.87 mL) and the resulting mixture was stirred at 90° C. for 1hour. The reaction was allowed to cool to room temperature andpartitioned between DCM (5 mL) and saturated aqueous sodium bicarbonate(5 mL). The layers were separated and the organic layer was concentratedin vacuo. Purification was by silica gel column chromatography elutingwith 0-50% ethyl acetate in heptane to give(6S,8R)-6-(5-bromopyridin-2-yl)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(89 mg, 55%) as a white solid. ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.16 (3H,d), 2.92-3.00 (2H, m), 3.25-3.36 (2H, m), 3.46-3.52 (1H, m), 5.07 (1H,s), 6.98 (1H, d), 7.09 (1H, dd), 7.43 (1H, d), 7.77 (1H, dd), 8.55 (1H,dd), 9.09 (1H, s); m/z: ES+ [M+H]+ 443.

Example 335-Fluoro-6-((6S,8R)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine

Sodium tert-butoxide (3.12 g, 32.52 mmol) and BrettPhos G3 (0.245 g,0.27 mmol) were added to a degassed solution of(6S,8R)-6-(5-bromo-3-fluoropyridin-2-yl)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(2.50 g, 5.42 mmol) and 1-(3-fluoropropyl)azetidin-3-amine (1.075 g,8.13 mmol) in 1,4-dioxane (27.1 mL) and reaction was warmed to 60° C.and stirred for 2 hours. After cooling, the reaction was diluted withEtOAc and water, and the layers were separated. The aqueous layer wasextracted with EtOAc, then the combined organics were dried over sodiumsulfate and evaporated. Purification was first by silica gel columnchromatography eluting with 0-100% (10% methanol in ethyl acetate) inheptane then by reverse phase Interchim (0.1% NH₃ and MeCN as eluents)to give the product as a mixture of isomers. The products were separatedby SFC; the sample was dissolved in MeOH and separated using thefollowing SFC conditions: Column: Phenomonex C4, 30×250 mm, 5 micron,mobile phase: 25% MeOH+0.1% NH₃, flow rate: 100 ml/min, BPR 120 bar,column temp: 40° C. to give5-fluoro-6-((6S,8R)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine(1.780 g, 64%) as a foam. ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.15 (3H, d),1.67-1.81 (2H, m), 2.59 (2H, t), 2.81-3.03 (4H, m), 3.17-3.34 (2H, m),3.70 (3H, q), 4.01-4.08 (1H, m), 4.20 (1H, d), 4.43 (1H, t), 4.53 (1H,t), 5.32 (1H, s), 6.54 (1H, dd), 6.81 (1H, d), 6.95 (1H, d), 7.64-7.68(1H, m), 9.46 (1H, s); m/z: ES+ [M+H]+ 513.

The(6S,8R)-6-(5-bromo-3-fluoropyridin-2-yl)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolineused as starting material was synthesised as follows:

(6S,8R)-6-(5-bromo-3-fluoropyridin-2-yl)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline

Trifluoroacetic acid (2.16 mL) was added to a solution of(R)-1-(3-fluoro-1H-indazol-4-yl)-N-(2,2,2-trifluoroethyl)propan-2-amine(2.50 g, 9.08 mmol) and 5-bromo-3-fluoropicolinaldehyde (1.85 g, 9.08mmol) in toluene (43.3 mL) and the resulting mixture was stirred at 90°C. for 90 minutes. The reaction was allowed to cool and partitionedbetween DCM (50 mL) and saturated aqueous sodium bicarbonate (50 mL).The layers were separated and the organic layer was concentrated invacuo. Purification was by silica gel column chromatography eluting with0-50% ethyl acetate in heptane to give(6S,8R)-6-(5-bromo-3-fluoropyridin-2-yl)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline(2.90 g, 69%) as a white solid. ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.15(3H, d), 2.88-3.04 (2H, m), 3.24-3.35 (2H, m), 3.63-3.71 (1H, m), 5.39(1H, s), 6.83 (1H, d), 7.08 (1H, dd), 7.60 (1H, dd), 8.36 (1H, dd), 9.13(1H, s); m/z: ES+ [M+H]+ 461.

Examples 34 to 73 (table below) were prepared using synthetic methodsanalogous to those described above.

LCMS Ex No Structure Name 1H NMR [M + H] 34

N-(1-(3- fluoropropyl)aze- tidin-3-yl)- N-methyl-6- ((6S,8R)-8-methyl-7- (2,2,2- trifluoroethyl)- 6,7,8,9- tetrahydro-3H- pyrazolo[4,3-f]isoquinolin- 6-yl)pyridin-3- amine ¹H NMR (500 MHz, CDCl₃, 27° C.)1.15 (3H, d), 1.65-1.86 (2H, m), 2.59 (2H, t), 2.74-2.84 (1H, m), 2.85(3H, s), 2.89- 3.09 (3H, m), 3.16- 3.41 (2H, m), 3.59 (1H, td),3.64-3.80 (2H, m), 4.10 (1H, q), 4.44 (1H, t), 4.53 (1H, t), 5.06 (1H,s), 6.89 (1H, d), 6.99 (1H, dd), 7.11 (1H, d), 7.30 (1H, d), 7.96- 7.98(2H, m), 10.78 (1H, s). 491 35

N-(1-(3- fluoropropyl)aze- tidin-3-yl)-6- ((6S,8R)-6- deuterio-8-methyl-7- (2,2,2- trifluoroethyl)- 6,7,8,9- tetrahydro-3H- pyrazolo[4,3-f]isoquinolin- 6-yl)pyridin-3- amine ¹H NMR (500 MHz, CDCl₃, 27° C.)1.14 (3H, d), 1.76 (2H, dq), 2.62 (2H, t), 2.85 (1H, dd), 2.93- 2.98(2H, m), 2.99- 3.04 (1H, m), 3.21- 3.31 (2H, m), 3.55- 3.63 (1H, m),3.73 (2H, s), 4.11 (2H, s), 4.44 (1H, t), 4.53 (1H, t), 6.80 (1H, dd),6.90 (1H, d), 7.16 (1H, d), 7.21 (1H, d), 7.84 (1H, d), 8.01 (1H, s) 47836

N-(1-(3- fluoropropyl)aze- tidin-3-yl)-6- ((6S,8R)-1- deuterio-8-methyl-7- (2,2,2- trifluoroethyl)- 6,7,8,9- tetrahydro-3H- pyrazolo[4,3-f]isoquinolin- 6-yl)pyridin-3- amine ¹H NMR (500 MHz, CDCl₃, 27° C.)1.14 (3H, d), 1.71-1.81 (2H, m), 2.62 (2H, t), 2.86 (1H, dd), 2.93- 3.04(3H, s), 3.20- 3.32 (2H, m), 3.59 (1H, d), 3.73 (2H, s), 4.11 (2H, s),4.43 (1H, t), 4.53 (1H, t), 5.03 (1H, s), 6.80 (1H, d), 6.90 (1H, d),7.18 (2H, dd), 7.84 (1H, s) 478 37

N-(1-(3- fluoropropyl)aze- tidin-3-yl)-5- methoxy-6- ((6S,8R)-8-methyl-7- (2,2,2- trifluoroethyl)- 6,7,8,9- tetrahydro-3H- pyrazolo[4,3-f]isoquinolin- 6-yl)pyridin-3- amine ¹H NMR (300 MHz, DMSO-d₆, 27° C.)1.08 (3H, d), 1.52- 1.78 (2H, m), 2.46 (2H, t), 2.63-2.84 (3H, m),2.87-3.07 (2H, m), 3.34-3.50 (1H, m), 3.58-3.69 (2H, m), 3.80 (3H, s),3.83-4.07 (2H, m), 4.45 (2H, dt), 5.42 (1H, s), 6.24 (1H, d), 6.54 (1H,d), 6.66 (1H, d), 7.17 (1H, d), 7.26 (1H, d), 8.03 (1H, s), 12.90 (1H,s). 507 38

5-((6S,8R)-1- fluoro-8- methyl-7- (2,2,2- trifluoroethyl)- 6,7,8,9-tetrahydro-3H- pyrazolo[4,3- f]isoquinolin- 6-yl)-N-(1-(3-fluoropropyl)aze- tidin-3- yl)pyrazin-2- amine ¹H NMR (500 MHz, CDCl₃,27° C.) 1.16 (3H, d), 1.68-1.85 (2H, m), 2.63 (2H, t), 2.83 (1H, dd),2.93- 2.98 (1H, m), 2.99- 3.07 (2H, m), 3.21 (1H, dd), 3.24-3.35 (1H,m), 3.49-3.54 (1H, m), 3.72 (2H, t), 4.44 (1H, t), 4.47 (1H, d), 4.53(1H, t), 5.05 (1H, s), 5.37 (1H, d), 6.89 (1H, d), 6.96 (1H, dd), 7.75(1H, d), 8.17 (1H, d), 10.63 (1H, s). 496 39

2-fluoro-6- ((6S,8R)-1- fluoro-8- methyl-7- (2,2,2- trifluoroethyl)-6,7,8,9- tetrahydro-3H- pyrazolo[4,3- f]isoquinolin- 6-yl)-N-(1-(3-fluoropropyl)aze- tidin-3- yl)pyridin-3- amine ¹H NMR (500 MHz, CDCl₃,27° C.) 1.14 (3H, d), 1.69-1.86 (2H, m), 2.63 (2H, t), 2.83 (1H, dd),2.91- 3.08 (3H, m), 3.13- 3.38 (2H, m), 3.52 (1H, ddd), 3.77 (2H, q),4.11 (1H, q), 4.31- 4.39 (1H, m), 4.44 (1H, t), 4.53 (1H, t), 4.95 (1H,s), 6.85 (1H, dd), 6.89-6.98 (2H, m), 7.23 (1H, d), 10.41 (1H, s) 513 40

6-fluoro-N-(1- (3- fluoropropyl)aze- tidin-3-yl)-3- ((6S,8R)-8-methyl-7- (2,2,2- trifluoroethyl)- 6,7,8,9- tetrahydro-3H- pyrazolo[4,3-f]isoquinolin- 6-yl)pyridin-2- amine ¹H NMR (500 MHz, CDCl₃, 27° C.)1.14 (3H, d), 1.83 (2H, dp), 2.76 (2H, s), 2.81-2.91 (1H, m), 2.90-3.03(1H, m), 3.11-3.32 (4H, m), 3.53 (1H, dt), 3.84 (2H, d), 4.46 (2H, q),4.55 (1H, t), 5.20 (1H, s), 5.98-6.10 (1H, m), 6.83 (1H, d), 6.99-7.08(1H, m), 7.23 (1H, d), 8.06 (1H, d), 10.48 (1H, s). 495 41

N-(2-fluoro-4- ((6R,8R)-8- methyl-7- (2,2,2,- trifluoroethyl)- 6.7,8,9-tetrahydro-3H- pyrazolo[4,3- f]isoquinolin- 6-yl)phenyl)-1- (3-fluoropropyl)aze- tidin-3- amine ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.14(3H, d), 1.69-1.83 (2H, m), 2.59 (2H, t), 2.77 (1H, dd), 2.84- 2.89 (2H,m), 2.89- 2.98 (1H, m), 3.03 (1H, dd), 3.21 (1H, dq), 3.38 (1H, ddd),3.71-3.80 (2H, m), 4.04-4.16 (2H, m), 4.44 (1H, t), 4.53 (1H, t), 4.97(1H, s), 6.41 (1H, t), 6.76 (1H, dd), 6.90 (1H, dd), 6.97 (1H, d), 7.28(1H, d), 8.06 (1H, d), 10.20 (1H, s). 494 42

N-(1-(3- fluoropropyl)aze- tidin-3-yl)-5- ((6S,8R)-8- methyl-7- (2,2,2-trifluoroethyl)- 6,7,8,9- tetrahydro-3H- pyrazolo[4,3- f]isoquinolin-6-yl)pyrazin-2- amine ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 1.11 (3H, d),1.49- 1.80 (2H, m), 2.42- 2.48 (2H, m), 2.76- 2.93 (3H, m), 2.93- 3.21(2H, m), 3.42- 3.69 (4H, m), 4.23- 4.34 (1H, m), 4.45 (2H, dt), 4.99(1H, s), 6.85 (1H, d), 7.25 (1H, d), 7.42 (1H, d), 7.78 (1H, d), 7.82(1H, d), 8.06 (1H, s), 12.98 (1H, s). 478 43

6-((6S,8R)-7- (2,2- difluoroethyl)- 8-methyl- 6,7,8,9- tetrahydro-3H-pyrazolo[4,3- f]isoquinolin- 6-yl)-N-(1-(3- fluoropropyl)aze-tidin-3-yl)- N- methylpyridin- 3-amine ¹H NMR (300 MHz, DMSO-d₆, 27° C.)1.06 (3H, d), 1.56- 1.78 (2H, m), 2.42- 2.48 (2H, m), 2.55- 2.70 (1H,m), 2.81 (3H, s), 2.82-2.95 (3H, m), 2.96-3.23 (2H, m), 3.39-3.54 (1H,m), 3.57-3.69 (2H, m), 4.05 (1H, quin), 4.45 (2H, dt), 4.93 (1H, s),5.88 (1H, tt), 6.77 (1H, d), 7.07 (2H, d), 7.21 (1H, d), 7.93 (1H, t),8.05 (1H, s), 12.95 (1H, s). 473 44

6-((6S,8R)-7- (2,2- difluoroethyl)- 6,8-dimethyl- 6,7,8,9-tetrahydro-3H- pyrazolo[4,3- f]isoquinolin- 6-yl)-N-(1-(3-fluoropropyl)aze- tidin-3- yl)pyridin-3- amine ¹H NMR (500 MHz, CDCl₃,27° C.) 1.20 (3H, d), 1.68-1.81 (3H, m), 1.87 (3H, s), 2.59 (2H, t),2.89 (2H, d), 3.01 (2H, dd), 3.32 (1H, dd), 3.54-3.62 (1H, m), 3.68-3.75(2H, m), 3.93 (1H, d), 4.07 (1H, q), 4.43 (1H, t), 4.53 (1H, t), 5.25-5.62 (1H, m), 6.68 (1H, dd), 7.01 (1H, d), 7.08 (1H, d), 7.20 (1H, d),7.83 (1H, d), 8.06 (1H, s), 10.01 (1H, s). 473 45

N-(4-((6S,8R)- 7-(2,2- difluoroethyl)- 6,8-dimethyl- 6,7,8,9-tetrahydro-3H- pyrazolo[4,3- f]isoquinolin- 6-yl)-3- methoxyphenyl)-1-(3- fluoropropyl)aze- tidin-3- amine ¹H NMR (500 MHz, CDCl₃, 27° C.)1.14 (3H, d), 1.68-1.82 (2H, m), 1.85 (3H, s), 2.58 (2H, t), 2.78- 2.99(5H, m), 3.17 (1H, dd), 3.44 (3H, s), 3.49-3.60 (1H, m), 3.66-3.78 (2H,m), 3.89 (1H, d), 4.03-4.14 (1H, m), 4.48 (2H, dt), 5.26- 5.54 (1H, m),5.97 (1H, dd), 6.03 (1H, d), 6.82 (1H, d), 7.00 (1H, d), 7.17 (1H, d),8.06 (1H, d), 9.95 (1H, s). 502 46

6-((6S,8R)-7- (2,2- difluoroethyl)- 8-methyl- 6,7,8,9- tetrahydro-3H-pyrazolo[4,3- f]isoquinolin- 6-yl)-5-fluoro- N-(1-(3- fluoropropyl)aze-tidin-3- yl)pyridin-3- amine ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.14 (3H,d), 1.66-1.82 (2H, m), 2.59 (2H, t), 2.72-2.80 (1H, m), 2.85 (1H, dd),2.90 (2H, d), 2.98-3.12 (1H, m), 3.24 (1H, dd), 3.49 (1H, d), 3.64-3.73(2H, m), 3.98-4.08 (1H, m), 4.16 (1H, d), 4.43 (1H, t), 4.53 (1H, t),5.28 (1H, s), 5.58- 5.89 (1H, m), 6.52 (1H, dd), 6.82 (1H, d), 7.19 (1H,d), 7.64- 7.69 (1H, m), 8.05 (1H, s), 10.04 (1H, 477 s). 47

6-((6S,8R)-7- (2,2- difluoroethyl)- 8-methyl- 6,7,8,9- tetrahydro-3H-pyrazolo[4,3- f]isoquinolin- 6-yl)-2-fluoro- N-(1-(3- fluoropropyl)aze-tidin-3- yl)pyridin-3- amine ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.12 (3H,d), 1.67-1.81 (2H, m), 2.59 (2H, t), 2.79 (1H, dddd), 2.86- 2.93 (2H,m), 3.04 (1H, ddd), 3.33 (1H, dd), 3.53-3.62 (1H, m), 3.69-3.79 (2H, m),4.06 (1H, q), 4.21 (1H, d), 4.44 (1H, t), 4.53 (1H, t), 4.89 (1H, s),5.48- 5.76 (1H, m), 6.75 (1H, dd), 6.93 (1H, d), 6.98 (1H, d), 7.19 (1H,d), 8.05 (1H, s), 9.99 (1H, s). 477 48

N-(4-((6R,8R)- 7-(2,2- difluoroethyl)- 8-methyl- 6,7,8,9- tetrahydro-3H-pyrazolo[4,3- f]isoquinolin- 6-yl)phenyl)-1- (3- fluoropropyl)aze-tidin-3- amine ¹H NMR (500 MHz, DMSO-d₆, 27° C.) 0.96 (3H, d), 1.52-1.64 (2H, m), 2.49- 2.61 (1H, m), 2.63- 2.78 (3H, m), 2.88- 3.02 (2H,m), 3.26- 3.31 (1H, m), 3.53- 3.64 (2H, m), 3.80- 3.87 (1H, m), 4.38(2H, dt), 4.74 (1H, s), 5.76 (1H, tt), 5.88- 5.93 (1H, m), 6.33 (2H, d),6.70 (1H, d), 6.80 (2H, d), 7.17 (1H, d), 7.98 (1H, s), 12.91 (1H, s).(Two hydrogens not observed). 458 49

N-(4-((6S,8R)- 7-(2,2- difluoroethyl)- 8-methyl- 6,7,8,9- tetrahydro-3H-pyrazolo[4,3- f]isoquinolin- 6-yl)-3,5- difluorophenyl)- 1-(3-fluoropropyl)aze- tidin-3- amine ¹H NMR (500 MHz, DMSO-d₆, 27° C.) 0.95(3H, d), 1.58- 1.70 (2H, m), 2.50- 2.70 (3H, m), 2.79 (1H, br dd), 2.86-2.97 (1H, m), 2.95- 3.12 (2H, m), 3.09 (1H, br dd), 3.33- 3.45 (1H, m),3.71- 3.88 (2H, m), 3.88- 4.01 (1H, m), 4.38 (2H, dt), 5.03 (1H, s),5.70 (1H, tt), 6.03 (2H, br d), 6.63 (1H, d), 6.69 (1H, br d), 7.13 (1H,d), 7.96 (1H, s), 12.89 (1H, s) 494 50

5-((6S,8R)-7- (2,2- difluoroethyl)- 8-methyl- 6,7,8,9- tetrahydro-3H-pyrazolo[4,3- f]isoquinolin- 6-yl)-N-(1-(3- fluoropropyl) azetidin-3-yl)pyrazin-2- amine ¹H NMR (500 MHz, DMSO-d₆, 27° C.) 1.06 (3H, d),1.56- 1.70 (2H, m), 2.42- 2.47 (2H, m), 2.57- 2.69 (1H, m), 2.75- 2.87(3H, m), 2.99- 3.13 (2H, m), 3.43- 3.51 (1H, m), 3.54- 3.62 (2H, m),4.29 (1H, sxt), 4.43 (2H, dt), 4.94 (1H, s), 5.97 (1H, tt), 6.79 (1H,d), 7.22 (1H, d), 7.41 (1H, d), 7.77 (1H, d), 7.84 (1H, d), 8.04 (1H,s), 12.97 (1H, s) 460 51

6-((6S,8S)-7- (2,2- difluoroethyl)- 8- (difluoromethyl)- 6,7,8,9-tetrahydro-3H- pyrazolo[4,3- f]isoquinolin- 6-yl)-N-(1-(3- fluoropropyl)azetidin-3- yl)pyridin-3- amine ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.68-1.86 (2H, m), 2.63 (2H, dd), 2.98 (2H, dd), 3.00-3.07 (1H, m), 3.20 (1H,dd), 3.28-3.44 (2H, m), 3.70 (3H, q), 4.10 (1H, dh), 4.43 (2H, t), 4.52(1H, t), 5.19 (1H, s), 5.43 (1H, tdd), 5.81 (1H, td), 6.77 (1H, dd),6.84 (1H, d), 7.09 (2H, t), 7.86 (1H, d), 8.00- 8.02 (1H, m). 495 52

N-(4-((6S,8R)- 7-((1- fluorocyclo- propyl)meth- yl)-8-methyl- 6,7,8,9-tetrahydro-3H- pyrazolo[4,3- f]isoquinolin- 6-yl)-3- methoxyphenyl)-1-(3- fluoropropyl)- N- methylazetidin- 3-amine ¹H NMR (500 MHz, CDCl₃,27° C.) 0.46- 0.62 (2H, m), 0.85- 1.03 (2H, m), 1.10 (3H, d), 1.66-1.89(2H, m), 2.56-2.62 (2H, m), 2.64 (1H, d), 2.80 (3H, s), 2.89 (1H, dd),2.98 (2H, dt), 3.09 (1H, dd), 3.33 (1H, dd), 3.73 (2H, q), 3.80 (1H, q),3.87 (3H, s), 4.08 (1H, p), 4.44 (1H, t), 4.53 (1H, t), 5.37 (1H, s),6.13 (1H, dd), 6.26 (1H, d), 6.79 (2H, t), 7.07 (1H, d), 8.05 (1H, s),510 10.53 (1H, s). 53

N-(2-fluoro-4- ((6S,8R)-7-((1- fluorocyclopro- pyl)methyl)-8- methyl-6,7,8,9- tetrahydro-3H- pyrazolo[4,3- f]isoquinolin- 6-yl)-5-methoxyphenyl)- 1-(3- fluoropropyl) azetidin-3- amine ¹H NMR (500 MHz,CDCl₃, 27° C.) 0.46- 0.52 (2H, m), 0.92- 1.01 (2H, m), 1.08 (3H, d),1.71-1.83 (2H, m), 2.55-2.64 (3H, m), 2.84-2.93 (3H, m), 3.02-3.11 (1H,m), 3.35-3.42 (1H, m), 3.77-3.89 (6H, m), 4.03-4.09 (1H, m), 4.12-4.18(1H, m), 4.50 (2H, dt), 5.30 (1H, s), 6.13 (1H, d), 6.65 (1H, d), 6.80(1H, d), 7.12 (1H, d), 8.05 (1H, s), 9.97 (1H, s). 514 54

5-((6S,8R)-7- ((1- fluorocyclopro- pyl)methyl)-8- methyl- 6,7,8,9-tetrahydro-3H- pyrazolo[4,3- f]isoquinolin- 6-yl)-N-(1-(3-fluoropropyl)aze- tidin-3- yl)pyridin-2- amine ¹H NMR (500 MHz, CDCl₃,27° C.) 0.47- 0.61 (2H, m), 1.04 (2H, d), 1.09 (3H, d), 1.66-1.81 (2H,m), 2.59 (2H, t), 2.72 (1H, dd), 2.84 (1H, dd), 2.92 (2H, td), 3.01 (1H,dd), 3.14 (1H, dd), 3.51-3.61 (1H, m), 3.68-3.83 (2H, m), 4.39 (1H, q),4.43 (1H, t), 4.52 (1H, t), 4.92 (1H, d), 4.96 (1H, s), 6.29 (1H, d),6.88 (1H, d), 7.17 (1H, d), 7.40 (1H, dd), 7.82 (1H, d), 8.04 (1H, d),467 11.07 (1H, s). 55

5-((6S,8R)-7- ((1- fluorocyclopro- pyl)methyl)-8- methyl- 6,7,8,9-tetrahydro-3H- pyrazolo[4,3- f]isoquinolin- 6-yl)-N-(1-(3-fluoropropyl)aze- tidin-3- yl)pyrazin-2- amine ¹H NMR (300 MHz, DMSO-d₆,27° C.) 0.47-0.63 (1H, m), 0.67-0.80 (1H, m), 0.81-1.01 (2H, m), 1.04(3H, d), 1.49- 1.78 (2H, m), 2.45 (2H, t), 2.57-2.94 (4H, m), 2.96-3.23(2H, m), 3.47-3.71 (3H, m), 4.22-4.34 (1H, m), 4.44 (2H, dt), 4.92 (1H,s), 6.82 (1H, d), 7.22 (1H, d), 7.35 (1H, d), 7.78 (1H, d), 7.82 (1H,d), 8.05 (1H, d), 12.94 (1H, br d). 468 56

N-(4-((6S,8R)- 7-((1- fluorocyclopro- pyl)methyl)-8- methyl- 6,7,8,9-tetrahydro-3H- pyrazolo[4,3- f]isoquinolin- 6-yl)-3- methoxyphenyl)-1-(3,3,3- trifluoropropyl) azetidin-3- amine ¹H NMR (500 MHz, CDCl₃, 27°C.) 0.48- 0.55 (2H, m), 0.90- 0.98 (2H, m), 1.09 (3H, d), 2.10-2.21 (2H,m), 2.57-2.72 (3H, m), 2.85-2.92 (3H, m), 3.07 (1H, dd), 3.34 (1H, dd),3.71-3.82 (3H, m), 3.86 (3H, s), 4.07- 4.14 (1H, m), 5.33 (1H, s), 5.96(1H, dd), 6.11 (1H, d), 6.75 (1H, d), 6.81 (1H, d), 7.11 (1H, d), 8.05(1H, d), 10.00 (1H, s). 532 57

6-((6S,8R)-7- ((1- fluorocyclopro- pyl)methyl)-8- methyl- 6,7,8,9-tetrahydro-3H- pyrazolo[4,3- f]isoquinolin- 6-yl)-N-(1-(3-fluoropropyl)aze- tidin-3-yl)- N- methylpyridin- 3-amine ¹H NMR (300MHz, DMSO-d₆, 27° C.) 0.40-0.57 (1H, m), 0.61-0.77 (1H, m), 0.78-1.00(2H, m), 1.02 (3H, d), 1.57- 1.75 (2H, m), 2.42- 2.48 (2H, m), 2.66 (1H,dd), 2.80 (3H, s), 2.83-2.93 (3H, m), 3.00 (1H, dd), 3.19 (1H, br dd),3.57-3.71 (3H, m), 4.04 (1H, quin), 4.45 (2H, dt), 4.88 (1H, s), 6.80(1H, d), 7.01- 7.10 (2H, m), 7.19 (1H, d), 7.93 (1H, d), 8.04 (1H, s),12.93 481 (1H, s). 58

2,2-difluoro-3- ((6S,8R)-6-(5- ((1-(3- fluoropropyl)aze- tidin-3-yl)oxy)pyridin- 2-yl)-8- methyl- 3,6,8,9- tetrahydro-7H- pyrazolo[4,3-f]isoquinolin- 7-yl)propan-1- ol ¹H NMR (500 MHz, DMSO, 27° C.) 1.06(3H, d), 1.65 (2H, dq), 2.61-2.71 (1H, m), 2.82 (1H, dd), 2.94 (2H, dd),3.00 (1H, dd), 3.11-3.21 (1H, m), 3.36-3.42 (1H, m), 3.64-3.74 (4H, m),4.39 (1H, t), 4.48 (1H, t), 4.80 (1H, p), 5.04 (1H, s), 5.39 (1H, t),6.84 (1H, d), 7.17-7.26 (3H, m), 8.02-8.07 (2H, m), 12.97 (1H, s). 49059

2,2-difluoro-3- ((6S,8R)-6-(3- fluoro-5-((1- (3- fluoropropyl)aze-tidin-3- yl)oxy)pyridin- 2-yl)-8- methyl- 3,6,8,9- tetrahydro-7H-pyrazolo[4,3- f]isoquinolin- 7-yl)propan-1- ol ¹H NMR (500 MHz, CDCl₃,27° C.) 1.17 (3H, d), 1.71-1.82 (2H, m), 2.65 (2H, t), 2.79-2.95 (3H,m), 3.11-3.18 (2H, m), 3.19-3.28 (1H, m), 3.33 (1H, dd), 3.68 (1H, td),3.84 (4H, ddt), 4.44 (1H, t), 4.53 (1H, t), 4.79 (1H, p), 5.39 (1H, s),6.75 (1H, d), 6.86 (1H, dd), 7.20 (1H, d), 7.94 (1H, d), 8.06 (1H, d).508 60

2,2-difluoro-3- ((6S,8R)-6-(4- ((1-(3- fluoropropyl)aze- tidin-3-yl)amino)-2- methoxyphenyl)- 8-methyl- 3,6,8,9- tetrahydro-7H-pyrazolo[4,3- f]isoquinolin- 7-yl)propan-1- ol ¹H NMR (500 MHz, CDCl₃,27° C.) 1.13 (3H, d), 1.39 (1H, h), 1.68-1.83 (2H, m), 2.59 (2H, t),2.82- 2.99 (4H, m), 3.13 (1H, dt), 3.24 (1H, dd), 3.30-3.39 (1H, m),3.56-3.86 (8H, m), 4.09 (1H, p), 4.44 (1H, t), 4.53 (1H, t), 5.28 (1H,s), 5.98 (1H, dd), 6.11 (1H, d), 6.59 (1H, d), 6.76 (1H, d), 7.16 (1H,d), 8.04 (1H, d). 518 61

2,2-difluoro-3- ((6S,8R)-1- fluoro-6-(5- ((1-(3- fluoropropyl)aze-tidin-3- yl)amino)pyridin- 2-yl)-8- methyl- 3,6,8,9- tetrahydro-7H-pyrazolo[4,3- f]isoquinolin- 7-yl)propan-1- ol 1H NMR (500 MHz, CDCl₃,27° C.) 1.15 (3H, d), 1.66-1.88 (2H, m), 2.62 (2H, t), 2.88 (1H, dd),2.92- 3.02 (3H, m), 3.14 (1H, dd), 3.22 (1H, td), 3.37 (1H, ddd),3.67-3.73 (2H, m), 3.76 (1H, td), 3.97- 4.15 (2H, m), 4.31 (1H, d), 4.44(1H, t), 4.53 (1H, t), 5.11 (1H, s), 6.72 (1H, dd), 6.79 (1H, d), 6.87(1H, d), 7.01 (1H, d), 7.93 (1H, d), 10.19 (1H, s). 507 62

2,2-difluoro-3- ((6S,8R)-1- fluoro-6-(6- fluoro-5-((1- (3-fluoropropyl)aze- tidin-3- yl)amino)pyridin- 2-yl)-8- methyl- 3,6,8,9-tetrahydro-7H- pyrazolo[4,3- f]isoquinolin- 7-yl)propan-1- ol ¹H NMR(500 MHz, CDCl₃, 27° C.) 1.17 (3H, d), 1.68-1.84 (2H, m), 2.62 (2H, t),2.88 (1H, dd), 2.92- 2.97 (3H, m), 3.08- 3.16 (1H, m), 3.16- 3.27 (1H,m), 3.35- 3.44 (1H, m), 3.71- 3.78 (2H, m), 3.83 (1H, td), 3.93-4.14(2H, m), 4.38 (1H, d), 4.44 (1H, t), 4.53 (1H, t), 5.03 (1H, s),6.59-6.79 (2H, m), 6.92 (1H, d), 7.09 (1H, dd), 10.06 (1H, s). 525 63

6-((6S,8R)-7- (2-fluoro-2- methylpropyl)- 8-methyl- 6,7,8,9-tetrahydro-3H- pyrazolo[4,3- f]isoquinolin- 6-yl)-N-(1-(3-fluoropropyl)aze- tidin-3- yl)pyridin-3- amine ¹H NMR (500 MHz, DMSO-d₆,27° C.) 0.94 (3H, d), 1.13- 1.27 (6H, m), 1.51- 1.67 (2H, m), 2.23- 2.34(1H, m), 2.61- 2.77 (4H, m), 2.97 (1H, br dd), 3.40- 3.48 (1H, m), 3.50-3.65 (2H, m), 3.81- 3.93 (1H, m), 4.38 (2H, dt), 4.80 (1H, s), 6.11 (1H,br d), 6.69-6.78 (2H, m), 6.95 (1H, d), 7.12 (1H, d), 7.66 (1H, d), 7.96(1H, s), 12.86 (1H, s). (Two hydrogens not 469 observed). 64

N-(4-((6S,8R)- 7-(2-fluoro-2- methylpropyl)- 8-methyl- 6,7,8,9-tetrahydro-3H- pyrazolo[4,3- f]isoquinolin- 6-yl)-3- methoxyphenyl)-1-(3- fluoropropyl)aze- tidin-3- amine ¹H NMR (500 MHz, CDCl₃, 27° C.)1.05 (3H, d), 1.26 (6H, dd), 1.74 (2H, ddd), 2.37 (1H, dd), 2.54- 2.64(2H, m), 2.71- 2.9 (4H, m), 3.26 (1H, d), 3.73 (3H, d), 3.84 (3H, s),3.93 (1H, s), 4.10 (1H, s), 4.43 (1H, t), 4.53 (1H, t), 5.30 (1H, s),5.94 (1H, dd), 6.11 (1H, d), 6.67 (1H, d), 6.80 (1H, d), 7.11 (1H, d),8.04 (1H, d), 10.06 (1H, s). 498 65

6-((6S,8R)-7- (2- fluoropropyl)- 8-methyl- 6,7,8,9- tetrahydro-3H-pyrazolo[4,3- f]isoquinolin- 6-yl)-N-(1-(3- fluoropropyl)aze- tidin-3-yl)pyridin-3- amine ¹H NMR (500 MHz, DMSO-d₆, 27° C.) 1.00 (3H, d), 1.22(3H, dd), 1.59-1.70 (2H, m), 2.65-2.86 (4H, m), 3.14 (1H, br dd), 3.48(1H, sxt), 3.59-3.73 (2H, br m), 3.94 (1H, sxt), 4.44 (2H, dt), 4.60(1H, br dsxt), 4.79 (1H, s), 6.19 (1H, br d), 6.74 (1H, d), 6.78 (1H,dd), 6.93 (1H, d), 7.16 (1H, d), 7.74 (1H, d), 8.03 (1H, s), 12.93 (1H,s). (Three hydrogen multiplet obscured by DMSO). 455 66

6-((6S,8R)-7- (2- fluoropropyl)- 8-methyl- 6,7,8,9- tetrahydro-3H-pyrazolo[4,3- f]isoquinolin- 6-yl)-N-(1-(3- fluoropropyl)aze- tidin-3-yl)pyridin-3- amine ¹H NMR (500 MHz, DMSO-d₆, 27° C.) 1.00 (3H, d), 1.13(3H, dd), 1.56-1.70 (2H, m), 2.28-2.41 (1H, m), 2.45 (2H, t), 2.72 (2H,q), 2.77- 2.87 (2H, m), 3.13 (1H, br dd), 3.46 (1H, sxt), 3.56-3.66 (2H,m), 3.92 (1H, sxt), 4.44 (2H, dt), 4.55-4.72 (1H, m), 4.77 (1H, s), 6.18(1H, d), 6.73 (1H, d), 6.79 (1H, dd), 6.93 (1H, d), 7.16 (1H, d), 7.75(1H, d), 8.02 (1H, s), 12.92 (1H, s) 455 67

1-(3- fluoropropyl)- N-(4-((6S,8R)- 7-(2- fluoropropyl)- 8-methyl-6,7,8,9- tetrahydro-3H- pyrazolo[4,3- f]isoquinolin- 6-yl)-3-methoxyphenyl) azetidin-3- amine ¹H NMR (400 MHz, DMSO-d₆, 27° C.) 1.01(3H, d), 1.29 (3H, dd), 1.56-1.76 (2H, m), 2.43-2.49 (1H, m), 2.57-2.66(1H, m), 2.69-2.89 (3H, m), 3.06 (1H, br dd), 3.35-3.42 (1H, m),3.63-3.74 (2H, m), 3.81 (3H, s), 3.89- 3.99 (1H, m), 4.45 (2H, dt),4.70-4.91 (1H, m), 5.17 (1H, s), 5.88 (1H, dd), 6.00 (1H, br d), 6.18(1H, d), 6.32 (1H, d), 6.69 (1H, d), 7.19 (1H, d), 8.03 (1H, s), 12.93(1H, s). (Two hydrogen multiplet 484 obscured by DMSO). 68

1-(3- fluoropropyl)- N-(4-((6S,8R)- 7-(2- fluoropropyl)- 8-methyl-6,7,8,9- tetrahydro-3H- pyrazolo[4,3- f]isoquinolin- 6-yl)-3-methoxyphenyl) azetidin-3- amine ¹H NMR (400 MHz, DMSO-d₆, 27° C.) 1.00(3H, d), 1.14 (3H, dd), 1.57-1.73 (2H, m), 2.20-2.33 (1H, m), 2.42-2.48(2H, m), 2.68-2.83 (4H, m), 3.11 (1H, br dd), 3.37-3.47 (1H, m),3.59-3.66 (2H, m), 3.81 (3H, s), 3.92 (1H, sxt), 4.45 (2H, dt),4.63-4.84 (1H, m), 5.14 (1H, s), 5.91 (1H, dd), 5.98 (1H, d), 6.18 (1H,d), 6.42 (1H, d), 6.65 (1H, d), 7.16 (1H, d), 8.02 (1H, s), 12.93 (1H,s) 484 69

N-(1-(3- fluoropropyl)aze- tidin-3-yl)-6- ((6S,8R)-7- isobutyl-8-methyl- 6,7,8,9- tetrahydro-3H- pyrazolo[4,3- f]isoquinolin-6-yl)pyridin-3- amine ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 0.73 (3H, d),0.81 (3H, d), 0.97 (3H, d), 1.51-1.81 (3H, m), 1.99 (1H, dd), 2.34 (1H,dd), 2.42-2.48 (2H, m), 2.74 (2H, td), 2.85 (1H, dd), 3.09-3.22 (1H, m),3.37-3.54 (1H, m), 3.56-3.71 (2H, m), 3.93 (1H, sxt), 4.45 (2H, dt),4.69 (1H, s), 6.12 (1H, d), 6.72- 6.84 (2H, m), 6.94 (1H, d), 7.16 (1H,d), 7.75 (1H, d), 8.03 (1H, s), 12.90 (1H, 451 s). 70

6-((6S,8R)-7- (2,2- difluoropropyl)- 8-methyl- 6,7,8,9- tetrahydro-3H-pyrazolo[4,3- f]isoquinolin- 6-yl)-N-(1-(3- fluoropropyl)aze- tidin-3-yl)pyridin-3- amine ¹H NMR (300 MHz, DMSO-d₆, 27° C.) 1.07 (3H, d), 1.58(3H, br t), 1.80-2.05 (2H, m), 2.53-2.72 (1H, m), 2.86 (1H, br dd),2.98-3.27 (3H, m), 3.43-3.54 (1H, m), 3.87 (1H, br s), 4.06 (1H, br d),4.22- 4.50 (3H, m), 4.52- 4.65 (2H, m), 4.99 (1H, br s), 6.56 (1H, brs), 6.80 (1H, d), 6.86-7.20 (2H, m), 7.24 (1H, brd), 7.73- 7.88 (1H, m),8.08 (1H, s), 9.54-10.04 (1H, m), 12.99 (1H, 473 br s) 71

5-fluoro-N-(1- (3- fluoropropyl)aze- tidin-3-yl)-6- ((6S,8R)-8-methyl-7- (2,2,3- trifluoropropyl)- 6,7,8,9- tetrahydro-3H-pyrazolo[4,3- f]isoquinolin- 6-yl)pyridin-3- amine ¹H NMR (500 MHz,DMSO, 27° C.) 1.04 (3H, d), 1.64 (2H, dq), 2.44 (2H, t), 2.66-2.84 (4H,m), 2.97 (1H, dd), 3.15- 3.26 (1H, m), 3.56- 3.64 (3H, m), 3.94 (1H, h),4.39 (1H, t), 4.48 (1H, t), 4.56- 4.82 (2H, m), 5.19 (1H, s), 6.61 (1H,d), 6.66-6.73 (2H, m), 7.20 (1H, d), 7.55 (1H, dd), 8.04 (1H, s), 12.95(1H, s). 509 72

(S)-6-(8,8- dimethyl-7- (2,2,2- trifluoroethyl)- 6,7,8,9- tetrahydro-3H-pyrazolo[4,3- f]isoquinolin- 6-yl)-N-(1-(3- fluoropropyl)aze- tidin-3-yl)pyridin-3- amine ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.06 (3H, s), 1.41(3H, s), 1.66-1.83 (2H, m), 2.61 (2H, t), 2.93 (3H, dt), 3.13-3.36 (2H,m), 3.36-3.56 (1H, m), 3.71 (2H, q), 4.10 (2H, d), 4.44 (1H, t), 4.53(1H, t), 4.98 (1H, s), 6.72 (1H, dd), 6.77 (1H, d), 6.98 (1H, d), 7.11(1H, d), 7.83 (1H, d), 8.04 (1H, d), 10.72 (1H, s). 491 73

(S)-6-(7-(2,2- difluoroethyl)- 8,8-dimethyl- 6,7,8,9- tetrahydro-3H-pyrazolo[4,3- f]isoquinolin- 6-yl)-N-(1-(3- fluoropropyl)aze- tidin-3-yl)pyridin-3- amine ¹H NMR (500 MHz, CDCl₃, 27° C.) 1.04 (3H, s), 1.41(3H, s), 2.85-2.95 (6H, m), 3.21 (2H, t), 3.21- 3.35 (2H, m), 3.52 (2H,t), 4.01 (2H, q), 4.44 (1H, t), 4.55 (1H, t), 4.82 (1H, dt), 4.96 (1H,s), 6.72 (1H, dd), 7.07 (1H, dd), 7.09-7.13 (2H, m), 8.04 (1H, s), 8.20(1H, s), 10.65 (1H, s). 473

The above description of illustrative embodiments is intended only toacquaint others skilled in the art with the Applicant's specification,its principles, and its practical application so that others skilled inthe art may readily adapt and apply the specification in its numerousforms, as they may be best suited to the requirements of a particularuse. This description and its specific examples, while indicatingembodiments of this specification, are intended for purposes ofillustration only. This specification, therefore, is not limited to theillustrative embodiments described in this specification, and may bevariously modified. In addition, it is to be appreciated that variousfeatures of the specification that are, for clarity reasons, describedin the context of separate embodiments, also may be combined to form asingle embodiment. Conversely, various features of the specificationthat are, for brevity reasons, described in the context of a singleembodiment, also may be combined to form sub-combinations thereof.

What is claimed is:
 1. A compound of Formula (I):

wherein: A is CR² or N; G is CR³ or N; D is CR⁴ or N; E is CR⁵ or N; Qis O, NH or NMe; R¹ is CH₂F, CHF₂ or CF₃; R² is H, F, Cl, Me, CN, OMe orOEt; R³ is H or F; R⁴ is H, F, CN or OMe; R⁵ is H or F; R⁶ is H, Me,CH₂F, CHF₂ or CF₃; R⁷ is H or Me; R⁸ is C₁₋₃ alkyl, CH₂F, CHF₂, CF₃ orC3-4 cycloalkyl; R⁹ is Me, F or CH₂F; R¹⁰ is Me, F, CH₂F, CHF₂, CF₃,CH₂OMe or CH₂OH; R¹¹ is H or F; or R¹⁰ and R¹¹ taken together with thecarbon atom to which they are attached form a cyclopropyl ring or anoxetane ring; R¹² is independently selected from F or Me; R¹³ is H or F;and a is 0, 1 or 2; or a pharmaceutically acceptable salt thereof.
 2. Acompound of Formula (I) or a pharmaceutically acceptable salt thereof,as claimed in claim 1, wherein Q is NH.
 3. A compound of Formula (I) ora pharmaceutically acceptable salt thereof, as claimed in claim 1,wherein Q is O.
 4. A compound of Formula (I) or a pharmaceuticallyacceptable salt thereof, as claimed in claim 1, wherein R⁶ is H.
 5. Acompound of Formula (I) or a pharmaceutically acceptable salt thereof,as claimed in claim 1, wherein R⁷ is H.
 6. A compound of Formula (I) ora pharmaceutically acceptable salt thereof, as claimed in claim 1,wherein the group —CH₂—C(R⁹)(R¹⁰)(R¹¹) in the compound of Formula (I) isselected from the group consisting of:


7. A compound of Formula (IA) as claimed in claim 1

wherein: Q is O, NH or NMe; R¹ is CH₂F, CHF₂ or CF₃; R⁶ is H or Me; R⁷is H or Me; R⁸ is Me, CHF₂ or cyclopropyl; R⁹ is Me or F; R¹⁰ is Me, F,CH₂F, CH₂OMe or CH₂OH; R¹¹ is H or F; or R¹⁰ and R¹¹ taken together withthe carbon atom to which they are attached form a cyclopropyl ring or anoxetane ring; R¹² is independently selected from H or Me; R¹³ is H or F;a is 0, 1 or 2; and Ring Y is selected from the group consisting of:

or a pharmaceutically acceptable salt thereof. 8-9. (canceled)
 10. Amethod for the prevention or treatment of cancer in a warm-bloodedanimal, in need of such treatment which comprises administering to saidanimal an effective amount of a compound of the Formula (I) or (IA) or apharmaceutically acceptable salt thereof, as claimed in any one ofclaims 1-7 or
 13. 11-12. (canceled)
 13. A compound of Formula (I) asclaimed in claim 1, wherein the compound is selected from the groupconsisting of:N-(4-((6S,8R)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;6-(((6S,8R)-7-(2,2-difluoro-3-methoxypropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;6-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;N-(4-((6S,8R)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;3-((6S,8R)-6-(2,6-difluoro-4-((1-(3-fluoropropyl)azetidin-3-yl)amino)phenyl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)-2,2-difluoropropan-1-ol;N-(4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;(6S,8R)-7-((1-fluorocyclopropyl)methyl)-6-(4-((1-(3-fluoropropyl)azetidin-3-yl)oxy)-2-methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline;N-(3,5-difluoro-4-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine;5-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)-4-methoxypyridin-2-amine;N-(4-((6S,8R)-7-((3-(fluoromethyl)oxetan-3-yl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;N-(3,5-difluoro-4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine;(6S,8R)-7-(2-fluoro-3-methoxy-2-methylpropyl)-6-(4-(1-(3-fluoropropyl)azetidin-3-yloxy)-2-methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline;N-(4-((6S,8R)-7-(2-fluoro-3-methoxy-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;2,2-difluoro-3-((6S,8R)-6-(5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol;5-fluoro-6-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;N-(4-((6S,8R)-7-(2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;N-(3-ethoxy-4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine;N-(4-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;(6S,8R)-7-(2,2-difluoroethyl)-6-(4-((1-(3-fluoropropyl)azetidin-3-yl)oxy)-2-methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline;3-((6S,8R)-6-(2,6-difluoro-4-(1-(3-fluoropropyl)azetidin-3-ylamino)phenyl)-8-methyl-8,9-dihydro-3H-pyrazolo[4,3-f]isoquinolin-7(6H)-yl)-2-fluoro-2-methylpropan-1-ol;6-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;(6S,8R)-7-(2,2-difluoroethyl)-6-(5-((1-(3-fluoropropyl)azetidin-3-yl)oxy)pyridin-2-yl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinoline;1-(3-fluoropropyl)-N-(4-((6R,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-amine;1-(3-fluoropropyl)-N-(3-methoxy-4-(((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)azetidin-3-amine;2-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine;5-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine;2,2-difluoro-3-((6S,8R)-6-(3-fluoro-5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol;6-((6S,8R)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,9,8-tetrahydro-3H-pyrazolo[4,3-f]isoquin-6-yl)-N-(3-fluoropropyl)azetidi-3-yl)pyridin-3-amine;5-fluoro-6-((6S,8R)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;N-(1-(3-fluoropropyl)azetidin-3-yl)-N-methyl-6-((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine;N-(1-(3-fluoropropyl)azetidin-3-yl)-5-methoxy-6-(((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine;5-((6S,8R)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyrazin-2-amine;2-fluoro-6-((6S,8R)-1-fluoro-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;6-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)-5-(((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-2-amine;N-(2-fluoro-4-((6R,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine;N-(1-(3-fluoropropyl)azetidin-3-yl)-5-(((6S,8R)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyrazin-2-amine;6-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)-N-methylpyridin-3-amine;6-((6S,8R)-7-(2,2-difluoroethyl)-6,8-dimethyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;N-(4-((6S,8R)-7-(2,2-difluoroethyl)-6,8-dimethyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;6-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-5-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;6-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-2-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;N-(4-((6R,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)phenyl)-1-(3-fluoropropyl)azetidin-3-amine;N-(4-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3,5-difluorophenyl)-1-(3-fluoropropyl)azetidin-3-amine;5-((6S,8R)-7-(2,2-difluoroethyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyrazin-2-amine;6-((6S,8S)-7-(2,2-difluoroethyl)-8-(difluoromethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;N-(4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)-N-methylazetidin-3-amine;N-(2-fluoro-4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-5-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;5-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-2-amine;5-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyrazin-2-amine;N-(4-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3,3,3-trifluoropropyl)azetidin-3-amine;6-((6S,8R)-7-((1-fluorocyclopropyl)methyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)-N-methylpyridin-3-amine;2,2-difluoro-3-((6S,8R)-6-(5-((1-(3-fluoropropyl)azetidin-3-yl)oxy)pyridin-2-yl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol;2,2-difluoro-3-((6S,8R)-6-(3-fluoro-5-((1-(3-fluoropropyl)azetidin-3-yl)oxy)pyridin-2-yl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol;2,2-difluoro-3-((6S,8R)-6-(4-((1-(3-fluoropropyl)azetidin-3-yl)amino)-2-methoxyphenyl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol;2,2-difluoro-3-((6S,8R)-1-fluoro-6-(5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol;2,2-difluoro-3-((6S,8R)-1-fluoro-6-(6-fluoro-5-((1-(3-fluoropropyl)azetidin-3-yl)amino)pyridin-2-yl)-8-methyl-3,6,8,9-tetrahydro-7H-pyrazolo[4,3-f]isoquinolin-7-yl)propan-1-ol;6-((6S,8R)-7-(2-fluoro-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;N-(4-((6S,8R)-7-(2-fluoro-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)-1-(3-fluoropropyl)azetidin-3-amine;6-((6S,8R)-7-(2-fluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;6-((6S,8R)-7-(2-fluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;1-(3-fluoropropyl)-N-(4-((6S,8R)-7-(2-fluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)azetidin-3-amine;1-(3-fluoropropyl)-N-(4-((6S,8R)-7-(2-fluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-3-methoxyphenyl)azetidin-3-amine;N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-7-isobutyl-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine;6-((6S,8R)-7-(2,2-difluoropropyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;5-fluoro-N-(1-(3-fluoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,3-trifluoropropyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)pyridin-3-amine;(S)-6-(8,8-dimethyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine;and(S)-6-(7-(2,2-difluoroethyl)-8,8-dimethyl-6,7,8,9-tetrahydro-3H-pyrazolo[4,3-f]isoquinolin-6-yl)-N-(1-(3-fluoropropyl)azetidin-3-yl)pyridin-3-amine,or a pharmaceutically acceptable salt thereof.
 14. A method according toclaim 10, wherein the cancer is breast or gynaecological cancer.
 15. Amethod according to claim 10, wherein the cancer is breast cancer.
 16. Amethod according to claim 10, wherein the cancer is ER+ve breast cancer.